Fulvestrant compositions and method of use

ABSTRACT

Provided are inclusion complexes comprising fulvestrant and a cyclodextrin. The complexes may be useful for treating various conditions, such as cancer and systemic lupus erythematosus. Also provided are methods of producing the inclusion complexes, methods of using the inclusion complexes in therapy, and kits and unit dosages comprising the complexes.

CROSS-REFERENCE TO RELATED APPLICATIONS

This patent application claims priority from U.S. 61/383,660, thecontent of which is incorporated herein by reference.

FIELD OF THE INVENTION

The present invention relates to inclusion complexes between fulvestrantand a cyclodextrin, and formulations thereof. Methods of preparingfulvestrant inclusion complexes are also provided. Moreover, the presentinvention relates to the use of inclusion complexes of, andpharmaceutical formulations comprising, fulvestrant and a cyclodextrinin the treatment of a disease or condition that is or is believed to beresponsive to anti-estrogen therapy and/or ER-downregulation, such ascancer.

BACKGROUND OF THE INVENTION

Fulvestrant (7-alpha-[9-(4,4,5,5,5-pentafluoropentylsulphinyl)nonyl]estra-1,3,5-(10)-trien-3,17-beta-diol), hasthe structural formula (I):

and is commercially available as an oily parenteral formulation formonthly intramuscular administration under the tradename FASTODEX®(AstraZeneca Pharmaceuticals LP). Fulvestrant contains 6 asymmetriccarbon atoms and a stereogenic sulphoxide in the side chain. The activeingredient of FASLODEX™ is a mixture of 2 diastereoisomers:fulvestrantsulphoxide A and B, having the same absolute configuration at each ofthe stereogenic centers in the steroid system but different absoluteconfigurations at the sulphur atom. Commercially available Fulvestrantis a mixture of two diastereoisomers, Fulvestrant Sulphoxide A andFulvestrant Sulphoxide B. Fulvestrant formulations have been describedin, for example, U.S. Pat. Nos. 6,774,122 and 7,456,160.

Fulvestrant acts as an estrogen receptor antagonist without agonistproperties, blocking the trophic actions of estrogens without itselfhaving any partial agonist (estrogen-like) activity on the endometrium.Fulvestrant binds to estrogen receptors (ERs) in a competitive mannerwith affinity comparable with that of estradiol and downregulates the ERprotein in human breast cancer cells. Data from pro-clinical studiesindicated that fulvestrant is effective against human breast cancercells and xenografts displaying acquired resistance to tamoxifen orletrozole. See, Osborne et al, Cancer Chemother. Pharmacol., 134(2):89-95 (1994); Osborne et al, Journal of the National Cancer Institute,87(10) 746-750 (1995); and Long and Jelovac, Clinical Cancer Research,8:2378-2388 (2002). FASLODEX® is currently indicated for use in thetreatment of hormone receptor positive metastatic breast cancer inpostmenopausal women with disease progression following antiestrogentherapy. Bross et al., The Oncologist, 7:477-480 (2002).

Fulvestrant's characteristics, such as having a very high lipophilicity,extremely low aqueous solubility and the fact that it is only ionized atvery high pH, present formulation challenges and account for itsadministration as an oily intramuscular injection. Commerciallyavailable FASLODEX® is an intramuscular injection of 250 mg fulvestrantin a sterile oily solution in either a single 5 ml pre-filled syringe ortwo 2.5 ml pre-filled syringes, as long acting injection(s). The longacting FASLODEX® intramuscular depot formulation contains benzyl alcoholand castor oil as solvents and is designed to deliver the dose of 250 mgof fulvestrant over a 1 month period from a single 5 ml intramuscularinjection in the buttock or two 2.5 ml intramuscular injection into thebuttocks. Bross et al., The Oncologist, 7:477-480 (2002). While thisrequired monthly dose of 250 mg or bi-monthly dose of 125 mg offulvestrant is effective, it takes approximately 3-6 months to achievesteady-state plasma levels of fulvestrant. See, e.g., Chia andGradishar, The Breast 17:S16-S21 (2008); and William et al., ClinicalBreast Cancer, 6(1):S23-S29 (2005). Decreasing the time in whichsteady-state plasma levels are reached following administration offulvestrant may reduce the time taken to achieve a therapeutic response,which can be particularly beneficial for patients who would otherwiseexperience disease progression early during endocrine treatmentRobertson, J. F. R, The Oncologist 12:744-784 (2007). In addition,increasing ER downregulation with higher doses of fulvestrant mayprovide a better treatment response. Id.

Alternative dosing regimens of FASLODEX® have been suggested as a way toenhance the efficacy of fulvestrant therapy. However, even ifalternative dosing regimens of FASLODEX® were to provide a therapeuticbenefit, administration of fulvestrant as an intramuscular injectionstill has various disadvantages. For example, injection-site reaction,including transient pain and inflammation, is one of the most commondrug-related events reported for FASLODEX®. Bross, The Oncologist,7:477-480 (2002). Other disadvantages associated with intramuscularadministration include nerve/bone damage during needle insertion, painand tissue damage, accidental injection of air into artery or vein,extreme pain and/or tissue damage. Such events would only increase ifnew dosing regimens require more frequent injections. Intramuscularinjections may also be inconvenient to the patient and be a source ofanxiety, which can adversely impact patient compliance. In one study ofpatient preference for administration of endocrine therapy by injectionor oral tablets, the majority of respondants generally preferredadministration via daily tablets. L. Fallowfield, et ah, Annals ofOncology, 17: 205-210 (2006). Given the importance of patient complianceto therapeutic outcome, patient preference is an importantconsideration. In addition, intramuscular injections are not suitablefor all patients. For example, patients with certain blood disorders(e.g., bleeding diatheses, thrombocytopenia) or receiving anticoagulantsmay not be suitable candidates for administration of fulvestrant byintramuscular injection.

Delivery of fulvestrant via non-invasive formulations, such as oraldelivery, have been explored, but adequate bioavailability, aqueoussolubility, and target formulation concentration could not be achieved,presumably due to the high lipophilicity and low aqueous solubility offulvestrant. See, e.g., Harrison M., et al., (2003) Proc. ASCO, 22: 45,abstract 311.

There therefore remains a need for additional fulvestrant formulations,such as formulations that render fulvestrant suitable for oral,intranasal and/or sublingual administration.

BRIEF SUMMARY OF THE INVENTION

The present invention provides inclusion complexes, kits, formulations,and unit dosages comprising fulvestrant and a cyclodextrin. Alsoprovided are methods of producing fulvestrant inclusion complexes andmethods of treating a disease or indication that is responsive tofulvestrant therapy comprising administering a fulvestrant inclusioncomplex or formulation thereof to an individual.

In one aspect, an inclusion complex comprising a) a cyclodextrin; and b)a compound of the formula (I):

or a salt thereof or a solvate of the foregoing, is provided. Thecompound of formula (I) is also referred to herein as “fulvestrant”. Insome embodiments of any of the aspects of the invention, the compound ofthe formula (I) is fulvestrant sulphoxide A, fulvestrant sulphoxide B,or a mixture of fulvestrant sulphoxide A and fulvestrant sulphoxide B.It is understood that fulvestrant salts, such as pharmaceuticallyacceptable salts, and solvates thereof, are also intended by thedescriptions provided herein. Thus, all salt and non-salt forms offulvestrant and solvates of the foregoing are embraced by the inventionand descriptions of fulvestrant provided herein. In some embodiments,the cyclodextrin is hydroxypropyl beta-cyclodextrin (HPBCD) or methylbeta-cyclodextrin (MBCD).

In some embodiments of any of the aspects of the invention, thecyclodextrin for use in the inclusion complexes and formulations hereinis a water soluble unsubstituted or substituted alpha-cyclodextrin(ACD), beta-cyclodextrin (BCD), or gamma-cyclodextrin (GCD). In someembodiments, the beta-cyclodextrin is selected from the group consistingof methyl beta-cyclodextrin (MBCD), hydroxypropyl beta-cyclodextrin(HPBCD), and sulfobutylether beta-cyclodextrin (SBEBCD). In someembodiments, the beta-cyclodextrin is methyl beta-cyclodextrin orhydroxypropyl beta-cyclodextrin. In some embodiments, thegamma-cyclodextrin is hydroxypropyl gamma-cyclodextrin (HPGCD). In onepreferred embodiment, the cyclodextrin is hydroxypropylbeta-cyclodextrin (HPBCD) or methyl beta-cyclodextrin (MBCD).

In one aspect, provided are methods for improving the solubility of acompound of the formula (I) in water comprising combining the compoundof the formula (I) with a cyclodextrin. In another aspect, a method ofincreasing the water solubility of a compound of formula (I) isprovided, wherein the method comprises forming an inclusion complex of acyclodextrin and a compound of the formula (I). In some embodiments, thesolubility of the compound of the formula (I), when present as aninclusion complex with a cyclodextrin in deionized water at 20° C., isincreased by at least about or about 1.5-fold or 2-fold, when comparedto the solubility of the compound of formula (I) in uncomplexed formunder the same conditions. In other embodiments, the solubility, such asthe aqueous solubility, of fulvestrant in an inclusion complex may beincreased by at least about or about any of 3, 4, 5, 10, 15, 20, 30, 40,50, 60, 70, 80, 90, 100, 200, 300, 400, 500, 1,000, 2,000-fold or moreover uncomplexed fulvestrant. Solubility comparisons may be assessed bymethods known to the skilled artisan, such as any of the specificmethods and conditions detailed herein. In one preferred embodiment, thecyclodextrin is hydroxypropyl beta-cyclodextrin (HPBCD) or methylbeta-cyclodextrin (MBCD).

A method of increasing the oral bioavailability of a compound of formula(I) is provided, wherein the method comprises forming an inclusioncomplex of a cyclodextrin and a compound of the formula (I). In someembodiments, the oral bioavailability of the compound of the formula(I), when present as an inclusion complex with a cyclodextrin, is atleast about 50% greater than the oral bioavailability of the compound ofthe formula (I) in uncomplexed form. Oral bioavailability, andcomparisons thereof, may be assessed by methods known in the art,including any of the specific methods described herein. In one preferredembodiment, the cyclodextrin is hydroxypropyl beta-cyclodextrin (HPBCD)or methyl beta-cyclodextrin (MBCD).

In some embodiments, an inclusion complex of a compound of formula (I)and a cyclodextrin are provided, wherein the inclusion complex iscapable of inducing a greater maximum concentration (Cm) of the compoundsystemically than what is achievable when the compound is administeredalone in the same amount and under the same conditions. In one aspect,an inclusion complex of the compound of formula (I) is capable ofinducing at least about or about 1.5 or 2 or more times greater C_(max)for the compound systemically than what is achievable when the compoundis administered alone in the same amount and under the same conditions.In some embodiments, the C_(max) of fulvestrant, when administered to anindividual as an inclusion complex with a cyclodextrin, is at leastabout 2 times greater than the C_(max) of fulvestrant administered aloneunder the same conditions. In one preferred embodiment, the cyclodextrinis hydroxypropyl beta-cyclodextrin (HPBCD) or methyl beta-cyclodextrin(MBCD).

In some embodiments, an inclusion complex of a compound of formula (I)and a cyclodextrin is provided, wherein the inclusion complex is capableof inducing a greater area under the plasma concentration vs. time curve(AUC) of the compound than what is achievable when the compound isadministered alone (in the absence of a cyclodextrin) in the same amountand under the same conditions. In one aspect, an inclusion complex ofthe compound of formula (I) is capable of inducing at least about orabout 1 or 2 or more times greater AUC of compound than what isachievable when the compound is administered alone in the same amountand under the same conditions. In some embodiments, the AUC offulvestrant, when administered to an individual as an inclusion complexwith a cyclodextrin, is at least about 2 times greater than the AUC offulvestrant administered alone in the same amount and under the sameconditions. In one preferred embodiment, the cyclodextrin ishydroxypropyl beta-cyclodextrin (HPBCD) or methyl beta-cyclodextrin(MBCD).

In some embodiments, an inclusion complex of a compound of formula (I)and a cyclodextrin is provided, wherein the inclusion complex is capableof inducing a change in the time to reach the maximum plasma level(T_(max)) of the compound than what is achievable when the compound isadministered alone (in the absence of a cyclodextrin) in the same amountand under the same conditions. In one aspect, T_(max) is shorter with afulvestrant inclusion complex. In one aspect, an inclusion complex ofthe compound of formula (I) is capable of reducing T_(max) by about 0.5or 1 or 2 fold over what is achievable when the compound is administeredalone in the same amount and under the same conditions. In someembodiments, the T_(max) of fulvestrant, when administered to anindividual as an inclusion complex with a cyclodextrin, is at leastabout 2 times lower than the T_(max) of fulvestrant administered alonein the same amount and under the same conditions. In one aspect, afulvestrant inclusion complex reduces T_(max) by at least about or aboutany of 1, 2, 3, 5, 10 and 12 hours or more. In one preferred embodiment,the cyclodextrin is hydroxypropyl beta-cyclodextrin (HPBCD) or methylbeta-cyclodextrin (MBCD).

In some embodiments, an orally, intranasally, sublingually, rectally, orvaginally administered inclusion complex of a compound of formula (I)and a cyclodextrin is provided, wherein the inclusion complex may beadministered together with a long acting intramuscular fulvestrantformulation (e.g., FASLODEX®), compensating for the time required forthe long acting intramuscular fulvestrant formulation alone to reachsteady state fulvestrant plasma concentrations. In one preferredembodiment, the cyclodextrin is hydroxypropyl beta-cyclodextrin (HPBCD)or methyl beta-cyclodextrin (MBCD).

In some embodiments, a composition comprising a cyclodextrin and acompound of the formula (I) are provided, wherein the molar ratio of thecompound of formula (I) to the cyclodextrin is from about 1:1 to about1:300. In one aspect, the composition comprises an inclusion complex ofa cyclodextrin and a compound of the formula (I). In another aspect, thecomposition comprises a physical mixture of a cyclodextrin and acompound of the formula (I), wherein the physical mixture does notcomprise or is substantially free of an inclusion complex of acyclodextrin and a compound of the formula (I). In one preferredembodiment, the cyclodextrin is hydroxypropyl beta-cyclodextrin (HPBCD)or methyl beta-cyclodextrin (MBCD).

In another aspect, a formulation comprising a) a cyclodextrin; b) acompound of the formula (I), or a salt thereof or solvate of theforegoing; and c) a carrier, are provided. In one aspect, theformulation comprises an inclusion complex of the compound and acyclodextrin. In one variation, the carrier is a pharmaceuticallyacceptable carrier. The carrier may be in liquid, solid or semi-solidform. When the carrier is a liquid, it may be aqueous or an organicsolvent, or a combination thereof in any amount. In one aspect, thecarrier is selected from the group consisting of a solvent, a complexingagent, a filler, a diluent, a granulating agent, a disintegrant, alubricant, a glidant, a pH-modifier, a tonicity modifier, an adjuvant, adye, a polymer-based film coating, and a binder. In some embodiments,the carrier is one or more of water for injection, microcrystallinecellulose, glucose, sodium lauryl sulphate, crosscarmellose sodium,colloidal silica, talc, magnesium stearate, sodium benzoate, aluminummagnesium silicate, lactose, methanol, ethanol, propanol, and acetone.More than one carrier may be employed and combinations of carriersprovided herein are intended. In one preferred embodiment, thecyclodextrin is hydroxypropyl beta-cyclodextrin (HPBCD) or methylbeta-cyclodextrin (MBCD).

Formulations comprising a cyclodextrin and fulvestrant may furthercomprise additional formulation components, also referred to herein asadditional agents. In some embodiments of the formulations describedherein, the formulation further comprises an antioxidant. In one aspect,the antioxidant is a water-soluble antioxidant. In another aspect, theantioxidant is fully or partially water-insoluble. When awater-insoluble antioxidant is employed, care may be taken to reduce oravoid displacement of fulvestrant in the inclusion complex by theantioxidant. For example, formulations comprising a fulvestrantinclusion complex and a water-insoluble antioxidant may contain arelatively small amount of the antioxidant or take other precautions toreduce or avoid adverse consequences of the antioxidant on an inclusioncomplex of fulvestrant and a cyclodextrin. In some embodiments, theformulation may include combinations of two or more of the antioxidantsas described herein. In some embodiments, the antioxidant is selectedfrom the group consisting of ascorbyl palmitate, butylatedhydroxyanisole, butylated hydroxytoluene, malic acid, propyl gallate,sodium bisulfite, sodium sulfite, sodium metabisulfite, potassiummetabisulfite, potassium bisulfite, sodium thiosulfate, sodiumformaldehyde sulfoxylate, L-ascorbic acid, D-ascorbic acid,acetylcysteine, cysteine, thioglycerol, thioglycoUic acid, thiolacticacid, thiourea, dithiothreitol, dithioeythreitol, glutathione,nordihydroguaiaretic acid, tocopherol, sodium ascorbate, hypophophorousacid, and fumaric acid. In some embodiments, the antioxidant is selectedfrom the group consisting of butylated hydroxyanisole, malic acid,sodium bisulfite, sodium sulfite, sodium metabisulfite, potassiummetabisulfite, potassium bisulfite, sodium thiosulfate, sodiumformaldehyde sulfoxylate, L-ascorbic acid, D-ascorbic acid, cysteine,thiolactic acid, glutathione, nordihydroguaiaretic acid, sodiumascorbate, hypophophorous acid and fumaric acid. In some embodiments,the antioxidant is a dietary antioxidant, including, but not limited to,vitamin E, vitamin C, beta-carotene, and selenium. Other additionalagents may be employed in the formulations. In one preferred embodiment,the cyclodextrin is hydroxypropyl beta-cyclodextrin (HPBCD) or methylbeta-cyclodextrin (MBCD).

In some embodiments, a formulation comprising (a) a cyclodextrin; (b) acompound of the formula (I), or a salt thereof, or solvate of theforegoing; and (c) a carrier, is a solid formulation. In someembodiments, the formulation is a semi-solid. In some embodiments, theformulation is a liquid. In one preferred embodiment, the cyclodextrinis hydroxypropyl beta-cyclodextrin (HPBCD) or methyl beta-cyclodextrin(MBCD).

Unit dosage forms of fulvestrant inclusion complexes and formulationscomprising fulvestrant and a cyclodextrin are also provided. In someembodiments, the compound of the formula (I) is present in an amount ofbetween about 0.1 mg and about 500 mg per unit solid or semi-soliddosage form or between about 0.1 mg/mL and about 50 mg/mL in a liquiddosage form. In some embodiments, the compound of the formula (I) ispresent in an amount of about 250 mg per unit solid or semi-solid dosageform or about 25 mg/mL in a liquid dosage form. In one preferredembodiment, the cyclodextrin is hydroxypropyl beta-cyclodextrin (HPBCD)or methyl beta-cyclodextrin (MBCD).

In another aspect, an inclusion complex described herein is in asubstantially pure form.

Methods of administering fulvestrant inclusion complexes andformulations to an individual are also provided, as are methods of usingsuch complexes and formulations in therapy. For example, methods oftreating a disease or condition that is or is believed to be responsiveto anti-estrogen therapy and/or ER-downregulation are provided, whereinthe methods comprise administering to an individual in need thereof afulvestrant inclusion complex or formulation comprising fulvestrant anda cyclodextrin, wherein fulvestrant is not complexed with thecyclodextrin. In one aspect, the disease or condition is cancer. Thecancer may be early stage cancer, non-metastatic cancer, primary cancer,advanced cancer, locally advanced cancer, metastatic cancer, cancer inremission, recurrent cancer, cancer in an adjuvant setting, cancer in aneoadjuvant setting, or cancer substantially refractory to hormonetherapy. In some embodiments, the cancer is selected from the groupconsisting of breast cancer, endometrial cancer, prostate cancer, andlung cancer. In one variation, the cancer is breast cancer, such ashormone receptor positive metastatic breast cancer. In another aspect,the disease or condition is systemic lupus erythematosus. In onepreferred embodiment, the cyclodextrin is hydroxypropylbeta-cyclodextrin (HPBCD) or methyl beta-cyclodextrin (MBCD).

In a particular aspect, a method of treating a cancer in an individualin need thereof is provided, wherein the method comprises administeringto the individual an effective amount of an inclusion complex of a) acyclodextrin; and b) a compound of the formula (I), or a salt thereof,or solvate of the foregoing. The method may also employ a fulvestrantformulation described herein, such as a formulation comprising a) acyclodextrin; b) a compound of the formula (I), or a salt thereof orsolvate of the foregoing, and c) a carrier (e.g., a pharmaceuticallyacceptable carrier). In one preferred embodiment, the cyclodextrin ishydroxypropyl beta-cyclodextrin (HPBCD) or methyl beta-cyclodextrin(MBCD).

In some embodiments, the individual of the methods provided herein is ahuman, and may be male or female. In some embodiments the human has oris suspected of having a disease or condition that is or is believed tobe responsive to anti-estrogen therapy and/or ER-downregulation, such ascancer. In some embodiments, the individual is a post-menopausal womanwho has or is suspected of having hormone receptor positive metastaticbreast cancer. In some embodiments, the individual is a post-menopausalwoman who has or is suspected of having hormone receptor positivemetastatic breast cancer with disease progression following hormone(e.g., anti-estrogen) therapy. In some embodiments the human has or issuspected of having systemic lupus erythematosus.

In one aspect of the methods, the fulvestrant inclusion complex, or afulvestrant formulation provided herein, is intranasally, orally,sublingually, rectally, and/or vaginally administered to the individualin need thereof. In some embodiments, the dosage of fulvestrant in theadministered dose is between about 0.1 mg and about 500 mg per unitsolid or semi-solid dosage form or between about 0.1 mg/mL and about 50mg/mL in a liquid dosage form.

In another aspect of the therapeutic methods, such as a method oftreating a cancer in an individual in need thereof, the individual isadministered a combination of an effective amount of a fulvestrantinclusion complex or formulation and one other pharmaceutical agent. Insome embodiments, the other pharmaceutical agent is an anti-canceragent. In some embodiments, the other pharmaceutical agent is selectedfrom the group consisting of anastrozole (e.g., ARIMIDEX®), letrozole(e.g., FEMARA®), exemestane (e.g., AROMASIN®), aminoglutethimide (e.g.,Cytadren®), vorozole (e.g., RIVIZOR®), formestane (e.g., LENTARON®),fadrozole (e.g., AFEMA®), testolactone (e.g., TESLAC®), gefitinib (e.g.,IRESSA®), trastuzumab (e.g., HERCEPTIN®), crlotimnib (e.g., TARCEVA®),tipifanib (e.g., ZARNESTRA®), uncomplexed fulvestrant (e.g., FASLODEX®(AstraZeneca Pharmaceuticals LP)), and an antioxidant. It is understoodthat in some variations of the methods provided herein, two or moreother pharmaceutical agents are intended for use in the methods, such astwo or more of the anti-cancer agents provided herein. In someembodiments, the inclusion complex and the other pharmaceutical agentare administered simultaneously or sequentially. In some embodiments,the inclusion complex and the other pharmaceutical agent areadministered concurrently. The inclusion complex and the otherpharmaceutical agent may be administered, whether simultaneously,sequentially or otherwise, via the same or different routes.

In one variation, the methods comprise administering a) a compositioncomprising and/or an inclusion complex of a cyclodextrin and a compoundof the formula (I), or a salt thereof or solvate of the foregoing, andb) a compound of the formula (I) in uncomplexed form. In a particularvariation, a composition comprising and/or an inclusion complex of acyclodextrin and a compound of the formula (I), or a salt thereof, orsolvate of the foregoing, is administered orally, intranasally,sublingually, rectally, and/or vaginally and a compound of the formula(I) is administered intramuscularly. In one preferred embodiment, thecyclodextrin is hydroxypropyl beta-cyclodextrin (HPBCD) or methylbeta-cyclodextrin (MBCD).

In another aspect, provided is the use of an inclusion complex of a) acyclodextrin; and b) a compound of the formula (I), or a salt thereof,or solvate of the foregoing, for the manufacture of a medicament for usein a method provided herein, such as a method of treating a cancer, suchas breast cancer, in an individual. In one preferred embodiment, thecyclodextrin is hydroxypropyl beta-cyclodextrin (HPBCD) or methylbeta-cyclodextrin (MBCD).

In another aspect, provided is a kit comprising an inclusion complex ofa) a cyclodextrin; and b) a compound of the formula (I), or a saltthereof or solvate of the foregoing, or a formulation thereof. The kitmay further comprise instructions for use, such as for use in thetreatment of a disease or condition that is or is believed to beresponsive to anti-estrogen therapy and/or ER-downregulation. In aparticular variation, a kit comprises instructions for use in thetreatment of cancer or systemic lupus erythematosus. The kit may alsocomprise another pharmaceutical agent. In one such aspect, a kit isprovided comprising a) a cyclodextrin; b) a compound of the formula (I),or a salt thereof, or solvate of the foregoing, or a formulationthereof; and c) an additional pharmaceutical agent selected fromanastrozole (e.g., ARIMIDEX®), letrozole (e.g., FEMARA®), exemestane(e.g., AROMASIN®), aminoglutethimide (e.g., Cytadren®), vorozole (e.g.,RIVIZOR®), formestane (e.g., LENTARON®), fadrozole (e.g., AFEMA®),testolactone (e.g., TESLAC®), gefitinib (e.g., IRESSA®), trastuzumab(e.g., HERCEPTIN®), edotinib (e.g., TARCEVA®), tipifarnib (e.g.,ZARNESTRA®), uncomplexed fulvestrant (e.g., FASLODEX® (AstraZenecaPharmaceuticals LP)), and an antioxidant. When uncomplexed fulvestrantis employed, in one aspect it is formulated in an oil-based carrier(e.g., arachis oil, sesame oil, castor oil, or neutral oil (e.g.,MIGLYOL® 810 or MIGLYOL® 812)), or in a non-aqueous ester solvent (e.g.,benzyl benzoate, ethyl oleate, isopropyl myristate, isopropyl palmitate,or a mixture or combination of any thereof) and an alcohol (e.g.,ethanol, benzyl alcohol, or a mixture or combination of any thereof). Inone preferred embodiment, the cyclodextrin is hydroxypropylbeta-cyclodextrin (HPBCD) or methyl beta-cyclodextrin (MBCD).

In another aspect, provided are methods of producing an inclusioncomplex of a compound of the formula (I) and a cyclodextrin, comprisingadmixing a compound of the formula (I), or salt thereof, or solvate ofthe foregoing, with a cyclodextrin. In some embodiments, the methodfurther comprises adding a solvent, mixed solvent, or buffer to thecompound of the formula (I), the cyclodextrin, and/or a mixture thereof.If a solvent, mixed solvent or buffer is employed to form an inclusioncomplex, in one aspect the solvent, mixed solvent or buffer is removedafter formulation of the inclusion complex to provide an inclusioncomplex free of or substantially free of solvent, mixed solvent orbuffer. In one preferred embodiment, the cyclodextrin is hydroxypropylbeta-cyclodextrin (HPBCD) or methyl beta-cyclodextrin (MBCD).

In another aspect, provided are methods of producing an inclusioncomplex of a cyclodextrin and a compound of the formula (I), or a saltthereof, or solvate of the foregoing, comprising the steps of a)admixing the compound of the formula (I) and the cyclodextrin; and b)adding a suitable amount of solvent, mixed solvent, or buffer to themixture of step (a) and mixing until a suspension or solution is formed.In some embodiments, the step (a) further comprises admixing a suitablepolymer. In some embodiments, the method utilizes a suitable amount ofsolvent, such as water for injection (WFI). In some embodiments, themethod utilizes a suitable amount of a buffer, which in one aspect is aphosphate-citrate buffer. In some embodiments, the method utilizes asuitable polymer, which in one aspect is selected frompolyvinylpyrrolidone, hydroxypropyl methylcellulose,carboxymethylcellulose, and PLASDONE® Povidone. In one preferredembodiment, the cyclodextrin is hydroxypropyl beta-cyclodextrin (HPBCD)or methyl beta-cyclodextrin (MBCD).

BRIEF DESCRIPTION OF THE FIGURES

FIG. 1 shows a fulvestrant HPLC calibration data plot.

FIG. 2 shows a fulvestrant aqueous solubility data plot in the presenceand absence of various concentrations of alpha-cyclodextrin (ACD).

FIG. 3 shows a fulvestrant aqueous solubility data plot in the presenceand absence of various concentrations of beta-cyclodextrin (BCD).

FIG. 4 shows a fulvestrant aqueous solubility data plot in the presenceand absence of various concentrations of methyl beta-cyclodextrin(MBCD).

FIG. 5 shows a fulvestrant aqueous solubility data plot in the presenceand absence of various concentrations of hydroxypropyl beta-cyclodextrin(HPBCD).

FIG. 6 shows a fulvestrant aqueous solubility data plot in the presenceand absence of various concentrations of sulfobutyletherbeta-cyclodextrin (SBEBCD).

FIG. 7 shows a fulvestrant aqueous solubility data plot in the presenceand absence of various concentrations of gamma-cyclodextrin (GCD).

FIG. 8 shows a fulvestrant aqueous solubility data plot in the presenceand absence of various concentrations of hydroxypropylgamma-cyclodextrin (HPGCD).

FIG. 9 shows a fulvestrant plasma concentration data plot generated forExperiment I (oral gavage of fulvestrant-HPBCD).

FIG. 10 shows a fulvestrant plasma concentration data plot generated forExperiment II (oral gavage of fulvestrant-MBCD).

FIG. 11 shows a fulvestrant plasma concentration data plot generated forExperiment II (oral gavage without cyclodextrin (CD)).

FIG. 12 shows average fulvestrant plasma concentration data plotgenerated for Experiments I, II, and III (oral gavage of HPBCD, MBCD orwithout CD).

FIG. 13 shows fulvestrant plasma concentration data plot generated fororal gavage of fulvestrant-HPBCD at the dose of 5 mg/kg.

FIG. 14 shows a plot of the mean fulvestrant plasma concentrations overtime.

FIG. 15 shows a simulation of blood concentrations resulting from oraldosing of fulvestrant-HPBCD 5 mg/kg 8-hourly to an “average dog”(Average concentration at steady state based on exponentials=26 ng/mL. Adotted line is drawn at the level of the purported therapeutic targetconcentration).

FIG. 16 shows a plot of Fulvestrant-HPBCD stability data (Batch 1_1stored at 25° C. and 40° C.; T=0 to T=24 months).

FIG. 17 shows a plot of Fulvestrant-HPBCD stability data (Batch 1_2stored at 25° C. and 40° C.; T=0 to T=24 months).

FIG. 18 shows a plot of Fulvestrant-HPBCD stability data (Batch 1_3stored at 2° C.; T=0 to T=18 months).

FIG. 19 shows a plot of Fulvestrant-HPBCD stability data (Batch 1_4stored at 2° C.; T=0 to T=18 months).

FIG. 20 shows a plot of Fulvestrant-RAMEB stability data (Batch 2_1stored at 2° C.; 25° C. and 40° C.; T=0 to T=18 months).

FIG. 21 shows a plot of Fulvestrant-RAMEB stability data (Batch 2_2stored at 2° C.; 25° C. and 40° C.; T=0 to T=18 months).

DETAILED DESCRIPTION OF THE INVENTION

The present invention provides inclusion complexes, kits, formulations,and unit dosages comprising fulvestrant and a cyclodextrin. Alsoprovided are methods of producing the inclusion complexes as well asmethods of treatment.

The inventors have discovered that inclusion complexes comprisingfulvestrant and a cyclodextrin strikingly enhance the aqueous solubilityof fulvestrant as compared to fulvestrant in an uncomplexed form,suggesting that such inclusion complexes enhance the oral, intranasaland/or sublingual bioavailability of fulvestrant and may find particularuse in administration of fulvestrant via these dosage routes.Fulvestrant inclusion complexes may also provide superior orcomplimentary pharmacokinetics as compared to uncomplexed fulvestrantdelivered in the same amount and under the same conditions, includinggreater C_(max) and AUC values and a reduction in T_(max). In addition,therapeutic plasma levels of fulvestrant may be achieved sooner with afulvestrant inclusion complex as compared to long acting intra-muscularinjectable formulations of uncomplexed fulvestrant, which can have aparticular benefit to patients.

Abbreviations and Definitions

Unless defined otherwise or clearly indicated by context, all technicaland scientific terms and abbreviations used herein have the same meaningas commonly understood by one of ordinary skill in the art to which thisinvention belongs.

As used herein, the term “inclusion complex” or “IC” is intended acomplex wherein a moiety of a compound (e.g., a moiety of fulvestrant)is inserted, at least partially, into the cavity of a cyclodextrin(e.g., methyl beta-cyclodextrin (MBCD) or hydroxypropylbeta-cyclodextrin (HPBCD)). The inserted compound of the inclusioncomplex is considered “complexed” with the cyclodextrin. A compound thatis not part of an inclusion complex is considered “alone” or“uncomplexed.”

As used herein, the term “solubility” intends the solubility withreference to the total amount of compound (e.g., including the amount ofcompound in both complexed and uncomplexed form).

As used herein, “treatment”, “treating”, or “treat” is an approach forobtaining beneficial or desired results, including clinical results.

With respect to a disease or condition, “delaying” means to defer,hinder, slow, retard, stabilize, and/or postpone development of one ormore symptoms of the disease or condition (e.g., cancer or systemiclupus erythematosus). This delay can be of varying lengths of time,depending on the history of the disease and/or individual being treated.As is evident to one skilled in the art, a sufficient or significantdelay can, in effect, encompass prevention, in that the individual doesnot develop the condition (e.g., cancer or systemic lupuserythematosus).

As used herein, “pharmaceutically acceptable” with respect to amaterial, refers to a material that is not biologically or otherwiseunsuitable, e.g., the material may be incorporated (e.g., at the time ofmanufacturing or administration) into a pharmaceutical compositionadministered to an individual without causing significant undesirablebiological effects. Pharmaceutically acceptable carriers or excipientshave preferably met the required standards of toxicological andmanufacturing testing and/or are included on the Inactive IngredientGuide prepared by the U.S. Food and Drug administration.

The term “effective amount” intends such amount of a compound (e.g.,fulvestrant) or an inclusion complex (e.g., an inclusion complexcomprising fulvestrant and a cyclodextrin), or a combination therapycomprising at least one of the foregoing, which should be effective in agiven therapeutic form. As is understood in the art, an effective amountmay be in one or more doses, i.e., a single dose or multiple doses maybe required to achieve the desired treatment endpoint. An effectiveamount may be considered in the context of administering one or moreadditional pharmaceutical agents in a combination therapy, and aninclusion complex may be considered to be given in an effective amountif, when administered sequentially, simultaneously, or continuously withone or more additional pharmaceutical agents, one or more desirable orbeneficial result(s) may be or are achieved.

“Combination therapy” intends a first therapy (e.g., fulvestrant or aninclusion complex thereof) used in conjunction with a second therapy(e.g., surgery and/or an additional pharmaceutical agent) for treating,stabilizing, preventing, and/or delaying a disease or condition.Administration in “conjunction with” another compound includesadministration in the same or different composition(s), eithersequentially, simultaneously, or continuously, through the same ordifferent routes within a given time period. In some embodiments, thecombination therapy optionally includes one or more pharmaceuticallyacceptable carriers or excipients, non-pharmaceutically activecompounds, and/or inert substances.

The term “simultaneous administration,” as used herein, means that afirst therapy and second therapy in a combination therapy areadministered with a time separation of no more than about 15 minutes,such as no more than about any of 10, 5, or 1 minutes. When the firstand second therapies are administered simultaneously, the first andsecond therapies may be contained in the same composition (e.g., acomposition comprising both a first and second therapy) or in separatecompositions (e.g., a first therapy in one composition and a secondtherapy is contained in another composition). When there is a timeseparation in the context of simultaneous administration (e.g., 5minutes), either the first therapy or the second therapy may beadministered first.

As used herein, the term “sequential administration” means that thefirst therapy and second therapy in a combination therapy areadministered with a time separation of more than about 15 minutes, suchas more than about any of 20, 30, 40, 50, 60, or more minutes. Eitherthe first therapy or the second therapy may be administered first. Thefirst and second therapies are contained in separate compositions, whichmay be contained in the same or different packages or kits.

As used herein, the term “concurrent administration” means that theadministration of the first therapy and that of a second therapy in acombination therapy overlap with each other.

Unless otherwise stated, “substantially pure” in reference to aninclusion complex intends a preparation of the inclusion complex thatcontains about or less than about 15% impurity, wherein the impurityintends a compound other than an inclusion complex of fulvestrant and acyclodextrin. Substantially pure preparations include preparations thatcontain less than about 15% impurity, such as preparations that containless than about any one of 15%, 12%, 10%, 8%, 5%, 3%, 2%, 1% and 0.5%impurity.

Reference to “about” a value or parameter herein includes (anddescribes) variations that are directed to that value or parameter perse. For example, a description referring to “about X” includes thedescription of “X”.

As used herein, the singular forms “a,” “or,” and “the” include pluralreferents unless the context clearly dictates otherwise.

It is understood that wherever embodiments are described herein with thelanguage “comprising,” otherwise analogous embodiments described interms of “consisting of and/or “consisting essentially of are alsoprovided.

Where aspects or embodiments of the invention are described in terms ofa Markush group or other grouping of alternatives, the present inventionencompasses not only the entire group listed as a whole, but each memberof the group individually and all possible subgroups of the main group,such as the main group absent one or more of the group members. Thepresent invention also envisages the explicit exclusion of one or moreof any of the group members.

Inclusion Complexes

Cyclodextrins (“CDs”) are cyclic oligosaccharide structures composed ofvarious glucopyranose units, such as 6, 7 and 8 units (alpha-, beta- andgamma-cyclodextrin, respectively) bound together to form a ring.Cyclodextrins characterized by a cone-like molecular shape andcomprising a relatively hydrophobic central cavity and hydrophilic outersurface. The hydrophobic nature of the central cavity, in certain cases,endows the cyclodextrin with the ability to form inclusion complexeswith a hydrophobic “guest” molecule. Polar and ionic groups aregenerally less likely to be included within the hydrophobic cavity thanless-polar and non-ionic groups. In these complexes, a guest molecule isheld within the hydrophobic cavity of the cyclodextrin “host” molecule.The inclusion complex may be stabilized by a number of forces, such asvan der Waals attractive forces, electrostatics and hydrogen bonding.

Although many factors affect the stability of an inclusion complex, theability of a cyclodextrin molecule to form an inclusion complex with aguest molecule is a function of two key factors. The fast is steric anddepends on the relative size of the cyclodextrin to the size of theguest molecule or certain key functional groups within the guest. Thesecond factor is the thermodynamic interactions among differentcomponents of the system, such as the cyclodextrin, the guest, and thesolvent, where present.

The binding of guest molecules within host cyclodextrins is not fixed orpermanent but rather is a dynamic equilibrium. See, e.g., Valle, ProcessBiochemistry. 2004, 39(9): 1033-46 and Loftsson et al., Journal ofPharmaceutical Sciences, 1996, 85(10): 1017-25. Inclusion of a guestmolecule in a cyclodextrin host may exert a profound effect on thephysicochemical properties of the guest molecule that is temporarilylocked or caged within the host cavity. Inclusion complexes may giverise to beneficial properties, which are not achievable otherwise, suchas: solubility enhancement of highly insoluble guests; stabilization oflabile guests against the degradative effects of oxidation, visible orUV light and heat; control of volatility and sublimation; physicalisolation of incompatible compounds; chromatographic separations; tastemodification by masking off flavors, unpleasant odors, and controlledrelease of drugs. Cyclodextrin inclusion complexes, in some cases, mayprovide favorable flow, binding, and compaction properties to aid indrug formulation (e.g., in facilitating tablet compression).

Importantly, however, the effect of inclusion of a guest molecule in acyclodextrin host remains unpredictable. For example, although variouscyclodextrin complexes have been reported to enhance the bioavailabilityof small molecule drugs, cyclodextrin inclusion complexes have also beenreported to have either no effect on host bioavailability or in factdecrease the bioavailability of certain guest compounds. R. Carrier, etal., Journal of Controlled Release, 123(2):78-99. The interaction ofcyclodextrins with labile compounds can also result in several outcomes:cyclodextrins can retard degradation, can have no effect on reactivity,or can accelerate drug degradation. Loftsson et al., supra. In addition,the unpredictability of thermodynamic quantities related to inclusioncomplex formation have also been reported. A. Steffen et al., ChemistryCentral Journal, 2007, 1:29.

Inclusion complexes of fulvestrant as provided herein were discovered tonotably enhance the aqueous solubility of fulvestrant as compared tofulvestrant in an uncomplexed form, suggesting that such inclusioncomplexes enhance the bioavailability of fulvestrant and may be used inthe oral, intranasal, and/or sublingual administration of fulvestrant.Administration of fulvestrant via non-parenteral routes is significantand may offer patients more convenient, non-invasive treatment options,reducing the incidence of adverse side effects associated withparenteral fulvestrant therapy, and enhancing overall patientcompliance. Non-parenteral fulvestrant formulations may also have apositive effect on overall therapeutic outcome. For example, suchformulations may be used alone, or together with parenteraladministration of fulvestrant, to achieve steady-state drug levelssooner than with presently available fulvestrant parenteralformulations. In addition, higher dosing levels of fulvestrant may beachievable with such formulations, which can enhance ER downregulationand may provide an additional therapeutic benefit.

Inclusion complexes provided herein comprise fulvestrant and acyclodextrin, or a salt thereof, or a solvate (e.g., hydrate) of theforegoing. The inclusion complexes may be used alone, or together withan additional pharmaceutical agent, in the treatment of a disease orcondition provided herein, such as cancer. Other agents may also beincorporated with the inclusion complex, where applicable. For example,an inclusion complex may comprise a cyclodextrin and fulvestrant, withan antioxidant that is not itself a pharmaceutical agent but providesother benefits, such as stabilizing the complex over time, and whichdoes not adversely effect the fulvestrant inclusion complex.Formulations comprising a fulvestrant inclusion complex, and optionaladditional pharmaceutical or other agents, together with a carrier, suchas a pharmaceutically acceptable aqueous carrier, are also provided.

In some embodiments, an inclusion complex is an inclusion complex of acyclodextrin and fulvestrant sulphoxide A, fulvestrant sulphoxide B, ora mixture of fulvestrant sulphoxide A and fulvestrant sulphoxide B. In aparticular variation, a preparation of an inclusion complex is apreparation of a cyclodextrin and substantially pure fulvestrantsulphoxide A or substantially pure fulvestrant sulphoxide B. In anothervariation, a preparation of an inclusion complex is a preparation of acyclodextrin and a mixture of fulvestrant sulphoxide A and fulvestrantsulphoxide B.

Examples of a cyclodextrin for use in the inclusion complex include, butare not limited to, water soluble unsubstituted or substitutedalpha-cyclodextrin (ACD), beta-cyclodextrin (BCD), andgamma-cyclodextrin (GCD). Examples of substituted beta-cyclodextrinsthat may be employed in the inclusion complexes herein include, but arenot limited to, methyl beta-cyclodextrin (MBCD), hydroxypropylbeta-cyclodextrin (HPBCD), and sulfobutylether beta-cyclodextrin(SBEBCD).). Examples of substituted gamma-cyclodextrins that may beemployed in the inclusion complexes herein include, but are not limitedto, hydroxypropyl gamma-cyclodextrin (HPGCD). Mixtures of cyclodextrinsmay also be employed. For example, a formulation comprising fulvestrantand a mixture of two or three or four or more cyclodextrins is alsoprovided. Where a more than one cyclodextrin is employed, it may be ofthe same cyclodextrin class (e.g., two alpha-cyclodextrins) or differentchemical classes (e.g., one alpha-cyclodextrin and one beta- orgamma-cyclodextrin). In one preferred embodiment, the cyclodextrin ishydroxypropyl beta-cyclodextrin (HPBCD) or methyl beta-cyclodextrin(MBCD).

In some embodiments, a cyclodextrin is obtained from a commercialsource, including, but not limited to cyclodextrins sold under thefollowing tradenames CAVASOL® W6 HP (Wacker Chemie AG, Munich, Germany),CAVASOL® W6 HP TL (Wacker Chemie AG, Munich, Germany), CAVAMAX® W6Pharma (Wacker Chemie AG, Munich, Germany), CAVASOL® W7 HP (WackerChemie AG, Munich, Germany), CAVASOL® W7 HP Pharma (Wacker Chemie AG,Munich, Germany), CAVASOL® W7 HP TL (Wacker Chemie AG, Munich, Germany),CAVASOL® W7 M (Wacker Chemie AG, Munich, Germany), CAVASOL® W7 M Pharma(Wacker Chemie AG, Munich, Germany), CAVASOL® W7 M TL (Wacker Chemie AG,Munich, Germany), CAVASOL® W8 HP (Wacker Chemie AG, Munich, Germany),CAVASOL® W8 HP Pharma (Wacker Chemie AG, Munich, Germany), KLEPTOSE® HPB(Roquette Pharma, Geneva, Ill.), and CAPTISOL® (Cydex Pharmaceuticals,Inc. Lenexa, Kans.). In some embodiments, the cyclodextrin is CAVASOL®W7 M Pharma. In some embodiments, the cyclodextrin is KLEPTOSE® HPB.

The inclusion complex may comprise fulvestrant or a salt thereof or asolvate (e.g. a hydrate or alcoholate) of the foregoing that ispartially or completely included into the cavity of a cyclodextrinmolecule. Accordingly, one or more cyclodextrin molecules may beassociated with each fulvestrant molecule. The complex may exist in avariety of molar ratios, which may be dependent on a variety of physicalfactors during the formation of the complex, and be transitional andvary during formation.

In some embodiments, fulvestrant is fully included into the cavity of acyclodextrin molecule. In some embodiments, fulvestrant is partiallyincluded into the cavity of a cyclodextrin molecule. In some embodimentsof the inclusion complex, the molar ratio of the fulvestrant tocyclodextrin is from any of about 1:1 to about 1:300; about 1:1 to about1:150; about 1:1 to about 1:100; about 1:1 to about 1:50, about 1:1 toabout 1:25; about 1:1 to about 1:10; about 1:1 to about 1:5; about 1:1to about 1:4; about 1:1 to about 1:3; about 1:1 to about 1:2.5; andabout 1:1 to about 1:2, or is about any of 1:1, 1:2, and 1:3.

The inclusion complexes described herein may increase the solubility offulvestrant. In one aspect, are provided methods for improving thesolubility of fulvestrant in water comprising complexing fulvestrantwith a cyclodextrin. In some embodiments, the solubility of fulvestrantin a fulvestrant inclusion complex, in deionized water at 20° C., is atleast about 10-fold greater than the solubility of fulvestrant inuncomplexed form. In some embodiments, a fulvestrant inclusion complexincreases the solubility of fulvestrant by at least any of about or byabout 10-, 25-, 50-, 75-, 100-, 250-, 500-, 750-, 1000-, 1500-, 2000-,3000-, or 4000-fold compared to the solubility of fulvestrant inuncomplexed form.

In some embodiments, the uncomplexed fulvestrant is present in anoil-based carrier (e.g., arachis oil, sesame oil, castor oil, or neutraloil (e.g., MIGLYOL™ 810 or MIGLYOL® 812)), or in a non-aqueous estersolvent (e.g., benzyl benzoate, ethyl oleate, isopropyl myristate,isopropyl palmitate, or a mixture or combination of any thereof) and analcohol (e.g., ethanol, benzyl alcohol, or a mixture or combination ofany thereof). In some embodiments, an uncomplexed fulvestrant isobtained from a commercial source, including, but not limited to, thefulvestrant formulation sold as the tradename FASLODEX®.

The inclusion complexes described herein may provide improvedpharmacokinetic properties for fulvestrant. Such changes inpharmacokinetic properties may result in desired therapeutic effects,such as a more rapid onset of therapeutic effect and/or less prolongedand/or reduced effects of a disease or condition for which fulvestranttreatment is obtained (e.g., cancer or systemic lupus erythematosus).

The inclusion complexes described herein may result in increased oralbioavailability of fulvestrant. In some embodiments, the oralbioavailability of fulvestrant from an inclusion complex comprisingfulvestrant and a cyclodextrin is at least any of about or about 2%, 5%,10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 60%, 70%, 80%, or 90%greater than the oral bioavailability of fulvestrant alone under thesame conditions. Methods of increasing oral bioavailability offulvestrant by at least any of about or by about any of 2%, 5%, 10%,15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 60%, 70%, 80%, or 90% ascompared to administration of uncomplexed fulvestrant under the sameconditions are provided, such methods comprising administeringfulvestrant in the form of an inclusion complex comprising fulvestrantand a cyclodextrin.

Bioavailability assessments and comparisons may be determined usingstandard techniques known in the art (e.g., measuring AUC(fulvestrantinclusion complex)/AUC(uncomplexed fulvestrant)×100). In some of theseembodiments, the conditions for assessing the amount of increase inbioavailability of an inclusion complex of fulvestrant as compared tofulvestrant in uncomplexed form include orally or intranasallyadministering any of about 5 mg/mL, about 10 mg/mL, about 15 mg/mL,about 20 mg/mL, about 25 mg/mL, about 30 mg/mL, about 35 mg/mL, about 40mg/mL, or about 50 mg/mL of fulvestrant (in the appropriate complexed oruncomplexed form) in a carrier and at room temperature. In someembodiments, the fulvestrant can be administered at room temperature inan aqueous solution (e.g., WFI) in the presence or absence of a buffer.

The inclusion complexes described herein may result in an increased Cmof fulvestrant following administration of an amount of fulvestrant asan inclusion complex when compared to administering the same amount offulvestrant, via the same route and under the same conditions, inuncomplexed form. The inclusion complexes described herein may alsoresult in an increased C_(max) of fulvestrant following administrationof an amount of fulvestrant as an inclusion complex when compared toadministering the same amount of fulvestrant via a differentadministration route, in uncomplexed form. For example, C_(max) offulvestrant may be increased upon oral and/or intranasal administrationof an amount of fulvestrant as an inclusion complex with a cyclodextrinwhen compared to administration of the same amount of uncomplexedfulvestrant as an intramuscular injection. In some embodiments, theinclusion complexes described herein may result in an increased C_(max)for fulvestrant within about 3 hours after oral administration of afulvestrant inclusion complex to an individual. In some embodiments, theC_(max) of fulvestrant from an inclusion complex comprising fulvestrantand a cyclodextrin is at least about or about any of 4, 5, 6, 7, 8, 9,10, 11, 12, 13, 15, 17, 19, 21, 23, 25, 27, 29, 31, 33, 35, 37, 40, 50,60, 70, 80, 90, or 100 times greater than the C_(max) of uncomplexedfulvestrant administered in the same amount and under the sameconditions as the inclusion complex. In some of these embodiments, theconditions comprise orally or intranasally administering any of about 5mg/mL, about 10 mg/mL, about 15 mg/mL, about 20 mg/mL, about 25 mg/mL,about 30 mg/mL, about 35 mg/mL, about 40 mg/mL, or about 50 mg/mL offulvestrant (in the appropriate complexed or uncomplexed form) at roomtemperature in an aqueous solution (e.g., WFI). Methods of increasingC_(max) in accordance with the descriptions herein are also provided.

The inclusion complexes described herein may result in a decreasedT_(max) for fulvestrant following administration. In some embodiments,the T_(max) of fulvestrant from an inclusion complex comprisingfulvestrant and a cyclodextrin is at least about or about any of 2%,5/%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 60%, 70%, 80%, or 90%lower than the T_(max) of fulvestrant administered alone under the sameconditions. The inclusion complexes described herein may also result ina decrease in T_(max) following administration of an amount offulvestrant as an inclusion complex when compared to administering thesame amount of fulvestrant via a different administration route, inuncomplexed form. For example, T_(max) of fulvestrant may be decreasedupon oral and/or intranasal administration of an amount of fulvestrantas an inclusion complex with a cyclodextrin when compared toadministration of the same amount of fulvestrant as an intramuscularinjection. The value of T_(max) upon oral and/or intranasaladministration of an amount of fulvestrant as an inclusion complex maybe at least about or about any of 2%, 5%, 10%, 15%, 20%, 25%, 30%, 35%,40%, 45%, 50%, 60%, 70%, 80%, or 90% lower than the T_(max) obtained byadministration of the same amount of uncomplexed fulvestrant as anintramuscular injection. In some of these embodiments, the conditionscomprise orally or intranasally administering any of about 5 mg/mL,about 10 mg/mL, about 15 mg/mL, about 20 mg/mL, about 25 mg/mL, about 30mg/mL, about 35 mg/mL, about 40 mg/mL, or about 50 mg/mL of fulvestrant(in the appropriate complexed or uncomplexed form) at room temperaturein an aqueous solution (e.g., WFI). Methods of decreasing T_(max) inaccordance with the descriptions herein are also provided.

The inclusion complexes described herein may result in decreasing thetherapeutic time of onset for fulvestrant such that the therapeuticeffect occurs sooner with a fulvestrant inclusion complex as compared tofulvestrant administered in an uncomplexed form. In one embodiment, thetherapeutic time of onset of fulvestrant from an inclusion complexcomprising fulvestrant and a cyclodextrin is decreased by at least aboutor about any of 2%, 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%,60%, 70%, 80%, or 90% than the therapeutic time of onset of uncomplexedfulvestrant administered in the same amount and under the sameconditions. In other embodiments an inclusion complex of fulvestrantdecreases the therapeutic time of onset for fulvestrant by at leastabout or about any of 2%, 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%,50%, 60%, 70%, 80%, or 90% as compared to the therapeutic time of onsetof uncomplexed fulvestrant administered in the same amount as anintramuscular injection. In one aspect, the fulvestrant inclusioncomplex is provided by oral or intranasal administration. In some ofthese embodiments, the conditions comprise orally or intranasallyadministering any of about 5 mg/mL, about 10 mg/mL, about 15 mg/mL,about 20 mg/mL, about 25 mg/mL, about 30 mg/mL, about 35 mg/mL, about 40mg/mL, or about 50 mg/mL of fulvestrant (in the appropriate complexed oruncomplexed form) at room temperature in an aqueous solution (e.g.,WFI). Methods of decreasing therapeutic time of onset for fulvestrant inaccordance with the descriptions herein are also provided.

In some embodiments, the inclusion complex comprising fulvestrant and acyclodextrin is in substantially pure form. In one variation, apreparation of substantially pure inclusion complex is provided whereinthe preparation contains about or less than about any one of 15/%, 12%,10%, 8%, 5%, 3%, 2%, 1% and 0.5% impurity.

The inclusion complexes described herein, formulations thereof, andmethods include all salt and solvate forms. For example, an inclusioncomplex may comprise a compound of formula (I) and/or a salt thereof. Aninclusion complex may also comprise a solvate of a compound of formula(I) or a salt thereof. Solvates may contain either stoichiometric ornon-stoichiametric amounts of a solvent, and are often formed during theprocess of crystallization. In one aspect, a solvent is water and thesolvate is a hydrate. In another aspect, a solvent is an alcohol and thesolvate is an alcoholate. In some embodiments, the inclusion complexesdescribed herein can exist in unsolvated forms as well as solvatedforms. The inclusion complexes may also include hydrated forms or saltforms.

Methods of Preparation

Also provided are methods of preparing the inclusion complexes describedherein. In some instances inclusion complexes may be prepared on thebasis of liquid state, solid state or semi-solid state reaction betweenthe components. This may be accomplished by dissolving the cyclodextrinand fulvestrant in a suitable solvent or mixture of solvents. Ifdesired, an inclusion complex so obtained may be subsequently isolatedby crystallization, evaporation, spray drying or freeze drying. In asolid state method, the two components may be screened to uniformparticle size and thoroughly mixed, at which point they may be ground ina high energy mill with optional heating, screening and homogenization.In a semi-solid state, the two components may be kneaded in the presenceof small amounts of a suitable solvent, and the complex so-formed, isoven dried, screened and homogenized. The liquid state reactiongenerally provides optimum conditions for completeness of reaction.

In one aspect is provided a method of producing an inclusion complexcomprising fulvestrant and a cyclodextrin by admixing fulvestrant with acyclodextrin. In some embodiments, the method further comprises adding asolvent, mixed solvent, or buffer to fulvestrant, the cyclodextrin,and/or mixture thereof.

In the preparation of a fulvestrant inclusion complex, the suitableamount of solvent, mixed solvent, or buffer may be added directly to asolid mixture of fulvestrant and the cyclodextrin. Alternatively, thesolvent, mixed solvent, or buffer may be added to either fulvestrant orthe cyclodextrin, and then added to the other of fulvestrant and thecyclodextrin. In some embodiments, the solvent, mixed solvent, or buffermay be added independently to each of fulvestrant and cyclodextrin,followed by combining fulvestrant and the cyclodextrin.

In one aspect, the method of producing an inclusion complex offulvestrant and a cyclodextrin comprises the steps of: (a) admixingfulvestrant and a cyclodextrin; and (b) adding a suitable amount ofsolvent, mixed solvent, or buffer to the mixture of step (a) and mixinguntil a suspension or solution is formed.

In some embodiments, a buffer is employed. For example, a formulationcomprising a fulvestrant inclusion complex for intranasal administrationin one aspect further comprises a buffer as detailed herein. Suitablebuffers include, without limitation, phosphate buffers (e.g.,phosphate-citrate), potassium hydrogen phthalate buffers, and acetatebuffers. In some embodiments, the buffer is a phosphate-citrate buffer.In some embodiments, the added buffer and/or resulting suspension orsolution has a pH between any of about 1.0, 2.0, and 3.0 and about 8.0,9.0 or 10.0; about 3.0 and about 8.0, about 4.0 and about 6.0, about 4.5and about 5.5; or a pH of greater than, less than, or about any of 1.0,2.0, 3.0, 3.5, 4.0, 4.5, 5.0, 5.5, 6.0, 7.0, 7.5, 8.0, 9.0, or 10.0. Inone aspect, a formulation comprising a fulvestrant inclusion complex forintranasal administration comprises a buffer and the formulation has apH of between about 5.0 and about 8.0. In some embodiments, a solvent isemployed. A solvent in one aspect is an organic solvent or water.Suitable organic solvents are known to those of skill in the art andinclude, but are not limited to, low chain alcohols (e.g. methanol;ethanol; i-propanol) and acetone. In some embodiments, the solvent is apolar solvent, such as water (e.g., ddH₂0), methanol, ethanol, andi-propanol. In some embodiments, a mixed solvent is employed. A mixedsolvent in one aspect comprises an organic solvent or water. In oneaspect, a mixed solvent is a mixture of water and an organic solvent.Suitable solvents, mixed solvents, or buffers include 100° % of ddH₂0,or ddH₂0 or buffer together with ethanol or methanol (1-99%). Variouscombinations of solvent, mixed solvent, or buffer for optimizing theproduction of an inclusion complex of fulvestrant and a cyclodextrin canbe determined by persons skilled in the art in view of the methodsdescribed herein.

For the methods of producing an inclusion complex of fulvestrant and acyclodextrin, fulvestrant may be admixed with the cyclodextrin at amolar ratio from about 0.2:1 to about 1:300. In some embodiments, themolar ratio is about 0.5:1 to about 1:150, or about 1:1 to about 1:75,or about 1:1 to about 1:50, or about 1:1 to about 125, or about 1:1 toabout 1:10, or about 1:1 to about 1:5, or about 1:1 to about 1:3, orabout 1:1 to about 1:2, or any of about 1:1, 1:2, 13, 1:4, 1:5, 1:6, and1:7.

In some embodiments of the methods of producing an inclusion complex,the solvent, mixed solvent, or buffer is heated to less than, greaterthan, or about any of 25° C., 30° C., 35° C., 40° C., 45° C., 50° C.,55° C., 60° C., 65° C., 75° C., or 80° C. (e.g., before, during and/orafter mixing). The solvent, mixed solvent, or buffer may be heated priorto and/or after being added to fulvestrant and/or cyclodextrin. In someembodiments, the solvent, mixed solvent, or buffer is heated greaterthan the preferred temperature for less than, greater than, or about anyof 0.1 hr, 0.2 hr, 0.3 hr, 0.5 hr, 0.75 hr, 1 hr, 2 hr, 3 hr, 4 hr, 5hr, 7 hr, 10 hr, 15 hr, 24 hr, 36 hr, or 48 hr.

During the formation of the inclusion complex between fulvestrant and acyclodextrin, a suitable polymer may be added which may enhance thesolubility and/or complexation ability of fulvestrant and cyclodextrininclusion complex. Accordingly, in some embodiments of the methods ofproducing an inclusion complex, a step of admixing fulvestrant and acyclodextrin further comprises admixing a suitable polymer. Suitablepolymers include, for example, polyvinylpyrrolidone, hydroxypropylmethylcellulose, carboxymethylcellulose, and PLASDONE® Povidone, andderivatives thereof. In some embodiments, the suitable polymer is awater-soluble polymer.

In some embodiments of the methods of producing an inclusion complex,the mixing is continued for at least any of about 0.1 hr, 02 hr, 0.3 hr,0.5 hr, 0.75 hr, 1 hr, 2 hr, 4 hr, 10 hr, 24 hr, 36 hr, or 48 hrfollowing formation of the suspension or solution. If heat is applied tothe solvent, mixed solvent, or buffer during a method of producing aninclusion complex, the described mixing of the components may occurprior to, simultaneously with, and/or after the application of saidheat.

In some embodiments, the method of producing an inclusion complexfurther comprises a step for drying the product obtained from the stepsof a) admixing fulvestrant and a cyclodextrin and b) adding a suitableamount of a solvent, mixed solvent or buffer to the mixture. In someembodiments, the drying comprises evaporation. In some embodiments, theevaporation occurs for greater than, less than, or about any of 0.1 hr,0.25 hr, 0.5 hr, 1 hr, 2 hr, 5 hr, 10 hr, 1 day, or 5 days. In someembodiments, the evaporation is conducted under vacuum (e.g., less thanany of about 0.75 atm, 0.5 atm, or 0.25 atm). In some embodiments, theevaporation is conducted under atmospheric pressure. In someembodiments, the drying comprises dry heat. In some embodiments, thedrying comprises spray-drying. In some embodiments, the drying comprisesfreeze-drying. In some embodiments, the drying comprisesspray-granulation.

In one aspect, is provided a method of producing an inclusion complexbetween fulvestrant and a cyclodextrin comprising the steps of: (a)mixing the appropriate amount of fulvestrant and cyclodextrin with orwithout a suitable polymer; (b) adding a suitable amount of solvent,mixed solvent, and/or buffer to the mixture of step (a) with vigorousmixing until a paste or a slurry is formed; (c) continuing the mixingwith further addition of solvent (e.g., water), mixed solvent, or bufferif necessary to maintain the paste or the slurry consistency, for asuitable period of time to form the inclusion complex; and (d) dryingthe product of step (c). In some embodiments of step (b), the buffer isa phosphate-citrate buffer and the pH is about 5. In some embodiments,the solvent added during steps (b) and (c) is heated. In someembodiments, wherein the solvent, mixed solvent, or buffer is deionizedwater and/or a buffer, the deionized water and/or a buffer is heated toabout 60° C. In some embodiments, the mixing is preferably continued fora period of time greater than 0.2 hours. In some embodiments, thevigorous mixing until a paste or a slurry is formed is conducted atabout 60° C.

In another aspect, is provided a method of producing an inclusioncomplex between fulvestrant and a cyclodextrin comprising the steps of(a) mixing suitable amounts of fulvestrant and a cyclodextrin with orwithout a suitable polymer; (b) adding of a suitable solvent, mixedsolvent, and/or buffer to the mixture of step (a) with mixing until aslurry, suspension or solution is formed; and (c) allowing the formationof the inclusion complex by evaporation of the water over a period oftime. In some embodiments of step (b), the buffer is a phosphate-citratebuffer and the buffer pH is about 5. In some embodiments, the solventadded during step (b) is heated. In some embodiments, wherein thesolvent, mixed solvent, or buffer is deionized water and/or a buffer,the deionized water and/or a buffer is heated to about 60° C. In someembodiments of step (c), heat is applied to increase the evaporationrate. In some embodiments, the evaporation is conducted at 40° C. Insome embodiment, evaporation in step (c) occurs for greater than about 1hour. In some embodiments, the evaporation is conducted under vacuum.

In another aspect, is provided a method of producing an inclusioncomplex between fulvestrant and a cyclodextrin comprising the steps of:(a) mixing suitable amounts of fulvestrant and a cyclodextrin with orwithout a suitable polymer, (b) adding of a suitable solvent, mixedsolvent, and/or buffer to the mixture of step (a) with mixing until aslurry, suspension or solution is formed; and (c) spray-drying theslurry, suspension or solution to obtain a solid drug-cyclodextrininclusion complex. In some embodiments, the buffer is aphosphate-citrate and the buffer pH is about 5. In some embodiments, thesuitable solvent, mixed solvent, and/or buffer added during step (b) isheated. In some embodiments, wherein the solvent, mixed solvent, orbuffer is deionized water and/or a buffer, the deionized water and/or abuffer is heated to about 60° C.

In another aspect, is provided a method of producing an inclusioncomplex between fulvestrant and a cyclodextrin comprising the steps of:(a) mixing suitable amounts of fulvestrant and a cyclodextrin with orwithout a suitable polymer, (b) adding of a suitable solvent, mixedsolvent, and/or buffer to the mixture of step (a) with mixing until asolution is formed; and (c) freeze-drying the solution to obtain a soliddrug-cyclodextrin inclusion complex. In some embodiments, the buffer isa phosphate-citrate buffer and the buffer pH is about 5. In someembodiments, the solvent, mixed solvent, and/or buffer added during step(b) is heated. In some embodiments, wherein the solvent, mixed solvent,or buffer is deionized water and/or a buffer, the deionized water and/ora buffer is heated to about 60° C.

In another aspect, is provided a method of producing an inclusioncomplex between fulvestrant and a cyclodextrin comprising the steps of(a) mixing suitable amounts of fulvestrant and a cyclodextrin with orwithout a suitable polymer, (b) adding of a suitable solvent, mixedsolvent, and/or buffer to the mixture of step (a) with mixing until aslurry, suspension or solution is formed; (c) adding inactivepharmaceutical excipients to the slurry, suspension or solution, withcontinued mixing and (d) spray-granulating the slurry, suspension orsolution to obtain solid particles, suitable for formulation into anoral formulation, containing a solid drug-cyclodextrin inclusioncomplex. In some embodiments, the buffer is a phosphate-citrate bufferand the buffer pH is about 5. In some embodiments, the solvent, mixedsolvent, or buffer added during step (b) may be heated. In someembodiments, wherein the solvent, mixed solvent, or buffer is deionizedwater and/or a buffer, the deionized water and/or a buffer is heated toabout 60° C. The inactive pharmaceutical excipients included to producean oral formulation according to step (c) may include commonly usedpharmaceutical excipients commonly used in the art, and/or thosedescribed herein.

In another aspect, is provided a method of producing an inclusioncomplex between fulvestrant and a cyclodextrin comprising the steps of:(a) mixing suitable amounts of fulvestrant and cyclodextrin with orwithout a suitable polymer; (b) adding a suitable solvent, mixedsolvent, and/or buffer to the mixture of step (a) with mixing until asolution is formed; (c) producing a liquid oral or intranasalformulation. Inactive pharmaceutical excipients may be added to thesolution, containing the liquid drug-cyclodextrin inclusion complex. Insome embodiments, the buffer is a phosphate-citrate buffer and thebuffer pH is about 5. In some embodiments, the solvent, mixed solvent,and/or buffer added during step (d) is heated. In some embodiments,wherein the solvent, mixed solvent, or buffer is deionized water and/ora buffer, the deionized water and/or a buffer is heated to about 60° C.The inactive pharmaceutical excipients included to produce a liquid oralor intranasal formulation according to step (c) may include commonlyused pharmaceutical excipients commonly used in the art, and/or thosedescribed herein.

In some embodiments of the methods for producing an inclusion complexbetween fulvestrant and a cyclodextrin described above, greater than anyof about 10%, 15%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 85%, 90%, 95%,97%, 98%, or 99% of fulvestrant is in complexed form.

A product obtained by any of the preceding processes is also providedfor herein.

Formulations

The inclusion complexes described herein (e.g., an inclusion complex offulvestrant with a cyclodextrin) may be used in the preparation of acomposition or formulation, such as a pharmaceutical composition orformulation, by combining an inclusion complex described with at leastone or any combination of more than one of a pharmaceutical acceptablecarrier, excipient, stabilizing agent and/or other agent, which areknown in the art, for use in the methods of treatment, methods ofadministration, and dosage regimes described herein. A pharmaceuticallyacceptable carrier may include, for example, solvents, mixed solvents,complexing agents, fillers, stabilizers, diluents, granulating agents,disintegrants, lubricants, glidants, pH-modifiers, tonicity modifiers,adjuvants, binders, etc., known to the skilled artisan that are suitablefor administration to an individual (e.g., a human). Combinations of twoor more carriers are also contemplated.

The pharmaceutically acceptable carrier(s) and any additionalcomponents, as described herein, should be compatible for use in theintended route of administration (e.g., oral, parenteral, intranasal, orsublingal) for a particular dosage form. Such suitability will berecognized by the skilled artisan, particularly in view of the teachingprovided herein. The formulations may vary or be tailored according tothe condition to be treated, the amount of compound to be administered,the condition of the individual, and other variables that will readilybe apparent to one of ordinary skill in the art in view of the teachingsprovided herein. The inclusion complexes may be formulated, for example,as solid, semi-solid, and liquid dosage forms, such as tablets, pills,powders, liquid solutions or suspensions, suppositories, injectable andinfusible solutions, and sprays. The preferred form depends on theintended mode of administration and therapeutic application. Thefollowing formulations, additives, and methods are merely exemplary andare in no way limiting.

Fulvestrant described herein may be formulated with a cyclodextrin andmay comprise one or more of the favorable properties described for theinclusion complexes herein (e.g., increased solubility). In someembodiments, fulvestrant is in the uncomplexed form in the presence of acyclodextrin. In some embodiments, is provided a mixture of fulvestrantin both complexed and uncomplexed form with a cyclodextrin (e.g., amolar ratio mixture of greater than, less than, or any of about 1:300,1:150, 1:75, 1:50, 1:25, 1:20, 1:15 1:10, 1:7.5, 1:5, 1:3, 12, 1:1, 2:1,3:1, 4:1, 5:1, 7.5:1, 10:1, 15:1, 25:1, 50:1, 100:1, 150:1 or 300:1 ofcomplexed fulvestrant to uncomplexed fulvestrant, respectively).Accordingly, in one aspect is provided a formulation comprisingfulvestrant, a cyclodextrin, and a carrier. In some embodiments, theformulation comprises an effective amount of fulvestrant, cyclodextrin,and a carrier. The formulation may comprise a molar ratio ofcyclodextrin to fulvestrant that is greater than, less than, or any ofabout 1:1, 2:1, 3:1, 4:1, 5:1, 7.5:1, 10:1, 15:1, 25:1, 50:1, 75:1,100:1, 150:1 or 300:1. Additionally, the formulation comprising thefulvestrant, a cyclodextrin, and a carrier may be further formulated inany manner described herein for the inclusion complex formulations, andmay be used in any of the methods described herein, as well as at anydosage described herein, for the inclusion complexes and/or inclusioncomplex formulations (e.g., to treat a condition, such as cancer orsystemic lupus erythematosus). These formulations also may provideimproved pharmacokinetic properties as described herein (e.g.,bioavailability, C_(max), T_(max), and time of onset) when compared touncomplexed fulvestrant administered under the same conditions.

In some embodiments, the formulation comprising fulvestrant and acyclodextrin, a complex of fulvestrant with a cyclodextrin, or a mixturethereof, is a sterile formulation.

Additives used with the inclusion complexes described herein (e.g., aninclusion complex of fulvestrant with a cyclodextrin) include, forexample, one or more excipients (e.g., one or more excipients),antioxidants (e.g., one or more antioxidants), stabilizers (e.g., one ormore stabilizers), preservatives (e.g., one or more preservatives,including antimicrobial preservatives), pH adjusting and bufferingagents (e.g., one or more pH adjusting and/or buffering agents),tonicity adjusting agents (e.g., one or more tonicity adjusting agents),thickening agents (e.g., one or more thickening agents), suspendingagents (e.g., one or more suspending agents), binding agents (e.g., oneor more binding agents), viscosity enhancing agents (e.g., one or moreviscosity enhancing agents), sweetening agent (e.g., one or moresweetening agent) and the like, either alone or together with one ormore additional pharmaceutical agents, provided that the additionalcomponents are pharmaceutically acceptable. In some embodiments, theformulation may include combinations of two or more of the additionalcomponents as described herein (e.g., any of 2, 3, 4, 5, 6, 7, 8, ormore additional components).

In some embodiments, antioxidants include, but are not limited to,ascorbyl palmitate, butylated hydroxyanisole, butylated hydroxytoluene,malic acid, propyl gallate, sodium bisulfite, sodium sulfite, sodiummetabisulfite, potassium metabisulfite, potassium bisulfite, sodiumthiosulfate, sodium formaldehyde sulfoxylate, L-ascorbic acid,D-ascorbic acid, acetylcysteine, cysteine, thioglycerol, thioglycoUicacid, thiolactic acid, thiourea, dithiothreitol, dithioerythreitol,glutathione, nordihydroguaiaretic acid, tocophcrol, sodium ascorbate,hypophophorous acid, and fumaric acid. In some embodiments, theantioxidant is selected from the group consisting of sodium bisulfite,sodium sulfite, sodium metabisulfite, potassium metabisulfite, potassiumbisulfite, sodium thiosulfate, sodium formaldehyde sulfoxylate,L-ascorbic acid, D-ascorbic acid, acetylcysteine, thioglycerol,thioglycoUic acid, thiolactic acid, thiourea, dithiothreitol,dithiocrythreitol, glutathione, nordihydroguaiaretic acid, sodiumascorbate and hypophophorous acid.

In some embodiments, the antioxidant is a dietary antioxidant,including, but not limited to, vitamin E, vitamin C, beta-carotene, andselenium.

In some embodiments, the additives include processing agents and drugdelivery modifiers and enhancers, such as, for example, calciumphosphate, magnesium stearate, talc, monosaccharides, disaccharides,starch, gelatin, cellulose, methyl cellulose, sodium carboxymethylcellulose, dextrose, polyvinylpyrrolidinone, low melting waxes, ionexchange resins, and the like, as well as combinations of any two ormore thereof. Other suitable pharmaceutically acceptable excipients aredescribed in REMINGTON'S PHARMACEUTICAL SCIENCES, Mack Pub. Co., NewJersey 18 edition (1996), HANDBOOK OF PHARMACEUTICAL EXCIPIENTS,Pharmaceutical Press and American Pharmacists Association, 5 edition(2006), and REMINGTON: THE SCIENCE AND PRACTICE OF PHARMACY, LippincottWilliams & Wilkins, Philadelphia, 20^(th) edition (2003) and 21^(st)edition (2005).

Formulations suitable for oral administration may comprise, for example,(a) liquid solutions, such as an effective amount of fulvestrant orcyclodextrin inclusion complex thereof dissolved in diluents, such aswater, saline, er other ingestable liquid such as orange juice, (b)capsules, sachets or tablets, each containing a predetermined amount offulvestrant or inclusion complex thereof, as solids or granules, (c)suspensions in an appropriate liquid, (d) suitable emulsions, and (e)powders. Tablet forms can include one or more of lactose, mannitol, cornstarch, potato starch, microcrystalline cellulose, acacia, gelatin,colloidal silicon dioxide, croscarmellose sodium, talc, magnesiumstearate, stearic acid, and other excipients, colorants, diluents,buffering agents, moistening agents, preservatives, flavoring agents,and pharmacologically compatible excipients. Lozenge forms can comprisefulvestrant or an inclusion complex thereof in a flavor, usually sucroseand acacia or tragacanth, as well as pastilles comprising fulvestrant oran inclusion complex thereof in an inert base, such as gelatin andglycerin, or sucrose and acacia, emulsions, gels, and the likecontaining, in addition to the active ingredient, such excipients as areknown in the art. Oral formulations may include any suitable dosage,including those described herein, such as any of about 0.1 mg, 1 mg, 5mg, 6 mg, 7 mg, 8 mg, 9 mg, 10 mg, 11 mg, 12 mg, 13 mg, 14 mg, 14 mg, 15mg, 16 mg, 17 mg, 18 mg, 19 mg, 20 mg, 25 mg, 30 mg, 35 mg, 40 mg, 45mg, 50 mg, 75 mg, 100 mg, 150 mg, 200 mg, 250 mg, 275 mg, 300 mg, 350mg, 400 mg, 450 mg, or 500 mg of fulvestrant per unit dose. As describedbelow, the formulations may be used for the treatment of a condition(e.g., cancer or systemic lupus erythematosus). Solid dosage forms fororal administration may be particularly useful for the treatment ofcancer.

The inclusion complexes can be enclosed in a hard or soft capsule, canbe compressed into tablets, or can be incorporated with beverages orfood or otherwise incorporated into the diet. Capsules can be formulatedby mixing the inclusion complex with an inert pharmaceutical diluent andinserting the mixture into a hard gelatin capsule of the appropriatesize. If soft capsules are desired, a slurry, suspension or solution ofthe inclusion complex may be encapsulated by machine into a gelatincapsule.

Liquid dosage forms for oral administration may include pharmaceuticallyacceptable solutions, suspensions, syrups, and elixirs containing inertdiluents commonly used in the art, such as water or buffers. Suchformulations may also comprise adjuvants, such as wetting, emulsifying,suspending, antioxidants, preservatives (e.g., antimicrobialpreservative), enhancing (e.g., viscosity-enhancing), sweetening,flavoring, and perfuming agents. Liquid dosage forms of the inclusioncomplexes for oral administration may be particularly useful for thetreatment of cancer.

It is understood that when a formulation comprising a fulvestrantcyclodextrin inclusion complex is employed, wherein the formulationfurther comprises an additional agent, which may be an additionalpharmaceutical or other agent, that the additional agent may be employedin a manner that reduces the number or severity of adverse effects thatthe additional agent may have on the fulvestrant cyclodextrin inclusioncomplex. For example, although an additional agent may confer a benefiton a formulation comprising a fulvestrant cyclodextrin inclusioncomplex, such an agent may also have the potential to displacefulvestrant in the inclusion complex or otherwise prevent or reduce thequality of fulvestrant inclusion. Such agents may still confer a benefitbut their adverse consequences may be reduced or avoided if used inreduced amounts. Alternatively, additional agents may be selected suchthat the additional agent or combination thereof is less likely toadversely affect the inclusion complex.

Formulations suitable for parenteral administration include aqueous andnon-aqueous, isotonic sterile injection solutions, which can containanti-oxidants, buffers, bacteriostats, and solutes that render theformulation compatible with the blood of the intended recipient, andaqueous and non-aqueous sterile suspensions that can include suspendingagents, solubilizers, thickening agents, stabilizing agents, andpreservatives. The formulations can be presented in unit-dose ormulti-dose sealed containers, such as ampules and vials, and can bestored in a freeze-dried (lyophilized) condition requiring only theaddition of the sterile liquid for reconstitution (e.g., water forinjection), immediately prior to use. Extemporaneous injection solutionsand suspensions can be prepared from sterile powders, granules, andtablets of the kind previously described.

Injectable preparations (for example, sterile injectable aqueous oroleaginous suspensions) may be formulated according to the known artusing suitable dispersing or wetting agents and suspending agents. Thesterile injectable preparation may also be a sterile injectable solutionor suspension in a nontoxic parenterally acceptable diluent or solvent,for example, as a solution in propylene glycol. The sterile injectablepreparation may also be a sterile powder to be reconstituted usingacceptable vehicles prior to administration. Among the acceptablevehicles and solvents that may be employed are water, Ringer's solution,and isotonic sodium chloride solution. In addition, sterile, fixed oilsmay be employed as a solvent or suspending medium. For this purpose anybland fixed oil may be employed including synthetic mono- ordiglycerides. In addition, fatty acids such as oleic acid may be used inthe preparation of injectables.

Formulation of the inclusion complex in liquid form (for oraladministration, parenteral administration, intranasal administration, orotherwise) may have a pH in the range of about 1.0 to about 10.0,including for example pH ranges of any of about 5.0 to about 8.0, about5.0 to about 7.0, about 5.0 to about 6.0, about 6.5 to about 7.5, andabout 6.5 to about 7.0. In some embodiments, the pH of the compositionis formulated to no less than about 6, 5, or 4, including for example apH of any of about 8, 7.5, 7, 6.5, 6, 5.5, 5, 4.5, or 4. The formulationcan also be made to be isotonic with blood by the addition of a suitabletonicity modifier, such as glycerol.

The inclusion complexes may also be formulated for administration byinhalation. Formulations suitable for aerosol administration whichcomprise the inclusion complex may include, for example, aqueous andnon-aqueous, isotonic sterile solutions, which can containanti-oxidants, buffers, bacteriostats, and solutes, as well as aqueousand non-aqueous sterile suspensions that can include suspending agents,solubilizers, thickening agents, stabilizing agents, and preservatives,alone or in combination with other suitable components, which can bemade into aerosol formulations to be administered via inhalation. Theseaerosol formulations can be placed into pressurized acceptablepropellants, such as dichlorodifluoromethane, propane, nitrogen, and thelike. They also can be formulated as pharmaceuticals for non-pressuredpreparations, such as in a nebulizer or an atomizer.

The inclusion complexes may also be formulated in the form ofsuppositories for rectal administration. These can be prepared by mixingthe agent with a suitable non-irritating excipient that is solid at roomtemperature but liquid at rectal temperature and therefore will melt inthe rectum to release the drug. Such materials include cocoa butter,beeswax and polyethylene glycols.

The inclusion complexes may also be formulated for topicaladministration, especially when the target of treatment includes areasor organs readily accessible by topical application, including diseasesof the eye, the skin, or the lower intestinal tract. Suitable topicalformulations are readily prepared for each of these areas or organs.

Topical application for the lower intestinal tract can be effected in arectal suppository formulation (see above) or in a suitable enemaformulation. Topically-applied transdermal patches may also be used.

Also provided are unit dosage forms comprising the inclusion complexesand formulations described herein. These unit dosage forms can be storedin a suitable packaging in single or multiple unit dosages and may alsobe further sterilized and sealed. For example, the pharmaceuticalformulation (e.g., a dosage or unit dosage form of a pharmaceuticalformulation) may include (i) a mixture of fulvestrant with acyclodextrin, and/or an inclusion complex thereof and (ii) apharmaceutically acceptable carrier. In some variations, the amount offulvestrant within the formulation is in any of the following ranges:about 0.1 to about 50 mg, about 1 to about 50 mg, about 5 to about 50mg, about 20 to about 50 mg, about 50 to about 100 mg, about 100 toabout 125 mg, about 125 to about 150 mg, about 150 to about 175 mg,about 175 to about 200 mg, about 200 to about 225 mg, about 225 to about250 mg, about 250 to about 300 mg, about 300 to about 350 mg, about 350to about 400 mg, about 400 to about 450 mg, about 450 to about 500 mg,about 500 mg to about 550 mg, about 550 mg to about 600 mg, about 600 mgto about 650 mg, about 650 mg to about 700 mg, about 700 mg to about 750mg, about 750 mg to about 800 mg. In some embodiments, the amount offulvestrant within the formulation (e.g., a dosage or unit dosage form)is in the range of about 1 mg to about 500 mg, such as any of about 5 mgto about 250 mg, about 20 mg to about 400 mg, or about 50 mg, 100 mg,150 m, 200 mg, 250 mg, 300 mg, 350 mg, 400 mg, 450 mg, 500 mg, 550 mg,600 mg, 650 mg, 700 mg, 750 mg, or 800 mg. In some of these embodiments,the formulation may comprise a molar ratio of a cyclodextrin tofulvestrant that is greater than, less than, or any of about 1:1, 2:1,3:1, 4:1, 5:1, 7.5:1, 10:1, 15:1, 25:1, 50:1, 75:1, 100:1, 150:1 or300:1. For example, in some embodiments wherein the amount of the amountof fulvestrant within the formulation is in the range of about 1 mg toabout 800 mg, such as any of about 5 mg to about 700 mg, 20 mg to about500 mg, or about 20 mg, 25 mg, 50 mg, 75 mg, 100 mg, 150 mg, 200 mg, 250mg, 300, 350 mg, 400 mg, 450 mg, 500 mg, 550 mg, 600 mg, 650 mg, 700 mg,750 mg, or 800 mg, the amount of a cyclodextrin in the formulation maybe any of about 5 mg to about 2500 mg, such as any of about 15 mg toabout 1500 mg, about 30 mg to about 750 mg, about 60 mg to about 300 mg,about 100 mg to about 200 mg, or about 60 mg, 75 mg, 100 mg, 125 mg, 150mg, 200 mg, 225 mg, 250 mg, 275 mg or 300 mg. In some embodiments, thecarrier is suitable for parental administration (e.g., intravenousadministration). In some embodiments, the carrier is suitable forintramuscular administration. In some embodiments, the carrier issuitable for oral administration. In some embodiments, the carrier issuitable for intranasal administration. In some embodiments, the carrieris suitable for sublingual, rectal or vaginal administration. In someembodiments, fulvestrant of the inclusion complex is the onlypharmaceutically active agent for the treatment of the condition (e.g.,cancer or systemic lupus erythematosus) that is contained in theformulation.

In some embodiments, are provided dosage forms (e.g., a unit dosageform) for the treatment of cancer or systemic lupus erythematosus,comprising (i) a mixture of a fulvestrant with a cyclodextrin, and/or aninclusion complex thereof wherein the amount of fulvestrant is in therange of about 0.1 mg to about 500 mg per unit dose, and (ii) apharmaceutically acceptable carrier. In some embodiments, the amount offulvestrant in the unit dosage form includes any of about 0.1 mg, 1 mg,5 mg, 6 mg, 7 mg, 8 mg, 9 mg, 10 mg, 11 mg, 12 mg, 13 mg, 14 mg, 14 mg,15 mg, 16 mg, 17 mg, 18 mg, 19 mg, 20 mg, 25 mg, 30 mg, 35 mg, 40 mg, 45mg, 50 mg, 75 mg, 100 mg, 150 mg, 200 mg, 250 mg, 275 mg, 300 mg, 350mg, 400 mg, 450 mg, 500 mg, 550 mg, 600 mg, 650 mg, 700 mg, 750 mg, or800 mg of fulvestrant.

In some embodiments, are provided formulations of the inclusioncomplexes described herein comprising one or more of a complexing agent,a filler, a diluent, a granulating agent, a disintegrant, a lubricant,or a glidant. The complexing agent, filler, diluent, granulating agent,disintegrant, lubricant, or glidant may be chosen from among theingredient listed in Table 1. In some embodiments, formulations maycontain zero, one, or more than one ingredient from each use category inTable 1. Formulations may additionally contain other complexing agents,fillers, diluents, granulating agents, disintegrants, lubricants, orglidants not listed in Table 1. Formulations may also contain additionalingredients that are not complexing agents, fillers, diluents,granulating agents, disintegrants, lubricants, or glidants.

TABLE 1 Use Ingredients Complexing agent Sodium hydrogen carbonate;ethanol; methanol Filler/diluent Microcrystalline cellulose; calciumcarbonate; glucose; calcium hydrogen phosphate; lactose; mannitol;sodium chloride; sucrose; dextrates; microfine cellulose; modifiedstarch; sucrose-dextrin co-precipitate Granulating agent Acaciamucilage; glucose; gelatine; povidone (PVP); starch mucilage; sucrose;tragacanth mucilage Disintegrant Sodium hydroxymethylcellulose; alginicacid; sodium alginate; aluminium magnesium silicate; carbon dioxide;carmellose sodium; cationic exchange resins; croscarmellose sodium;microcrystalline cellulose; modified starch; sodium lauryl sulphate;sodium glycine carbonate; sodium starch glycollate; starch LubricantMagnesium stearate; calcium stearate; stearic acid; fumaric acid;hydrogenated vegetable oil; liquid paraffin; magnesium lauryl sulphate;macrogol 4000 and 6000; sodium benzoate; sodium lauryl sulphate; sodiumstearyl fumarate Glidant Colloidal silica; talc

Exemplary formulations are shown in Table 2.

TABLE 2 Formulation 1 Formulation 2 Formulation 3 Formulation 4Formulation 5 Complexing Sodium Ethanol None Methanol None agenthydrogen carbonate Filler/diluent Glucose Mannitol; MicrocrystallineSodium Glucose; sucrose modified starch cellulose; lactose chloride;dextrates Granulating None Acacia mucilage None Starch mucilageTragacanth agent mucilage Disintegrant Alginic acid; AluminumCrosscarmellose Microcrystalline Sodium lauryl carbon dioxide magnesiumsodium cellulose sulphate silicate Lubricant Calcium stearate Fumaricacid Magnesium Liquid paraffin Sodium stearate benzoate GlidantColloidal silica Talc Colloidal silica Talc Talc Other Dye Dye Dye;PVA-based PVA-based film None ingredients film coating coating

In some embodiments, the complexing agent, filler, diluent, granulatingagent, disintegrant, lubricant, or glidant is present in the amount pertablet indicated in Table 3.

TABLE 3 Amount per Ingredient tablet Complexing agent 1-200 mgFiller/diluent 100-200 mg Granulating agent 1-50 mg Disintegrant 1-50 mgLubricant 1-10 mg Glidant 5-30 mg

In some embodiments, the inclusion complex is formulated as a tabletcomprising fulvestrant and a cyclodextrin in amounts per tablet asindicated in Table 4 and one or more additional ingredients listed inTable 4 in an amount per tablet as indicated in Table 4.

TABLE 4 Ingredient Amount Per Tablet fulvestrant 0.1 mg, 1 mg, 5 mg, 6mg, 7 mg, 8 mg, 9 mg, 10 mg, 11 mg, 12 mg, 13 mg, 14 mg, 14 mg, 15 mg,16 mg, 17 mg, 18 mg, 19 mg, 20 mg, 25 mg, 30 mg, 35 mg, 40 mg, 45 mg, 50mg, 75 mg, 100 mg, 150 mg, 200 mg, 250 mg, 275 mg, 300 mg, 350 mg, 400mg, 450 mg, 500 mg, 550 mg, 600 mg, 650 mg, 700 mg, 750 mg, or 800 mgcyclodextrin 150, 160, 170, 180, 190, 200, 210, 220, 230, 240, 250, 275,300, 350, 400, 450, 500, 550, 600, 650, 700, 750, or 800 mgMicrocrystalline cellulose, glucose, mannitol, 150, 160, 165, 170, 180,185, 190, 195, or 200 mg or sucrose Aluminum magnesium silicate,carmellose 5, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, or 60 mg sodium,crosscarmellose, or modified starch Colloidal silica or talc 1, 2, 3, 4,5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20 mgDextrates, lactose, or sodium chloride 0.5, 1.0, 1.5, 2.0, 2.5, 3.0,3.5, 4.0, 4.5, or 5.0 mg Dye 0.5, 1.0, 1.5, 2.0, 2.5, 3.0, 3.5, 4.0,4.5, or 5.0 mg Polymer-based film coating 1, 2, 3, 4, 5, 6, 7, 8, 9, 10,11, 12, 13, 14, 15, 16, 17, 18, 19, or 20 mg

In some embodiments, are provided formulations of the inclusioncomplexes described herein comprising one or more of a vehicle, asuspending agent, antimicrobial preservatives, antioxidants, viscosityenhancing agents, sweetening agents, flavoring agents or coloringagents. The vehicle, suspending agent, antimicrobial preservatives,antioxidants, viscosity enhancing agents, sweetening agents, flavoringagents or coloring agents may be chosen from among the ingredientslisted in Table 5. In some embodiments, formulations may contain zero,one, or more than one ingredient from each use category in Table 5.Formulations may additionally contain another vehicle, a suspendingagent, antimicrobial preservatives, antioxidants, viscosity enhancingagents, sweetening agents, flavoring agents or coloring agents notlisted in Table 5. Formulations may also contain additional ingredientsthat are not vehicles, suspending agents, antimicrobial preservatives,antioxidants, viscosity enhancing agents, sweetening agents, flavoringagents, or coloring agents. In some embodiments, the formulationsdescribed herein are oral formulations.

TABLE 5 Use Ingredients Vehicle Water, distilled water, purified water,water for injection, aromatic water, juices, syrups, spirits Suspendingagent Agar, alginic acid, bentonite, calcium stearate, carbomers,carboxymethylcellulose calcium, carboxymethylcellulose sodium,carrageenan, cellulose, ceratonia, colloidal silicon dioxide, dextrin,gelatin, guar gum, hydroxyethyl cellulose, hydroxyethylmethyl cellulose,hydroxypropyl cellulose, hypromellose, kaolin, magnesium aluminumsilicate, maltitol, medium-chain triglycerides, methylcellulose,microcrystalline cellulose, polycarbophil, polyethylene glycol,potassium alginate, povidone, propylene glycol alginate, sesame oil,sodium alginate, sodium starch glycolate, sorbitan esters, sucrose,tragacanth, xanthan gum. Antimicrobial Alcohol, benzalkonium chloride,benzethonium chloride, benzoic acid, preservative benzyl alcohol, boricacid, bronopol, butylated hydroxyanisole, butylparaben, cetrimide,cetylpyridinium chloride, chlorbutanol, chlorhexidine, chlorobutanol,chlorocresol, chloroform, chloroxylenol, cresol, dimethyl ether,ethylparaben, glycerin, hexetidine, imidurea, isopropyl alcohol, lacticacid, methylparaben, monothioglycerol, parabens, alkyl chain length,phenol, phenoxyethanol, phenylethyl alcohol, phenylmercuric acetate,phenylmercuric borate, phenylmercuric nitrate, potassium benzoate,potassium metabisulfite, potassium sorbate, propionic acid, propylgallate, propylene glycol, propylparaben, sodium acetate, sodiumbenzoate, sodium borate, sodium lactate, sodium metabisulfite, sodiumpropionate, sodium sulfite, sorbic acid, thimerosal, xylitol.Antioxidant Ascorbyl palmitate, butylated hydroxyanisole, butylatedhydroxytoluene, malic acid, propyl gallate, sodium bisulfite, sodiumsulfite, sodium metabisulfite, potassium metabisulfite, potassiumbisulfite, sodium thiosulfate, sodium formaldehyde sulfoxylate,L-ascorbic acid, D-ascorbic acid, acetylcysteine, cysteine,thioglycerol, thioglycoUic acid, thiolactic acid, thiourea,dithiothreitol, dithioerythreitol, glutathione, nordihydroguaiareticacid, tocopherol, sodium ascorbate, hypophophorous acid, fumaric acid.Viscosity Acacia, agar, alginic acid, bentonite, carbomers,carboxymethylcellulose enhancing agent calcium, carboxymethylcellulosesodium, carrageenan, ceratonia, cetostearyl alcohol, chitosan, colloidalsilicon dioxide, cyclomethicone, ethylcellulose, gelatin, glycerin,glyceryl behenate, guar gum, hectorite, hydrogenated vegetable oil typeI, hydroxyethyl cellulose, hydroxyethylmethyl cellulose, hydroxypropylcellulose, hydroxypropyl starch, hypromellose, magnesium aluminumsilicate, maltodextrin, methylcellulose, polydextrose, polyethyleneglycol, poly(methylyinyl ether/maleic anhydride), polyvinyl acetatephthalate, polyvinyl alcohol, potassium chloride, povidone, propyleneglycol alginate, saponite, sodium alginate, sodium chloride, stearylalcohol, sucrose, tragacanth, xanthan gum Sweetening agent Acesulfamepotassium, alitame, aspartame, dextrose, erythritol, fructose, glycerin,inulin, isomalt, lactitol, glucose, maltitol, maltitol, maltose,mannitol, neohesperidin dihydrochalcone, polydextrose, saccharin,saccharin sodium, sodium cyclamate, sorbitol, sucralose, sucrose,syrups, honey, thaumatin, trehalose, xylitol.

Exemplary formulations (e.g., oral formulation) are shown in Table 6.

TABLE 6 Formulation 1 Formulation 2 Formulation 3 Formulation 4Formulation 5 Vehicle Water for Aromatic water Syrup Water for Water forinjection injection injection Suspending None None None NoneMicrocrystalline agent cellulose Antimicrobial None Benzoic acidChloroform Benzoic acid Benzoic acid preservative Antioxidant NoneThioglycerol Acetyl cysteine Thioglycerol Ascorbic acid Viscosity NoneNone None None Carboxymethyl enhancing cellulose agent SweeteningAspartame Acesulfame Fructose Saccharin Aspartame agent potassium OtherFlavouring & Flavouring & Flavouring & Flavouring & Flavouring &ingredients coloring agent coloring agent coloring agent coloring agentcoloring agent

In some embodiments, are provided formulations of the inclusioncomplexes described herein comprising one or more of a vehicle,suspending agents, antimicrobial preservatives, antioxidants, viscosityenhancing agents. The vehicle, suspending agents, antimicrobialpreservatives, antioxidants, viscosity enhancing agents may be chosenfrom among the ingredients listed in Table 7. In some embodiments,formulations may contain zero, one, or more than one ingredient fromeach use category in Table 7. Formulations may additionally containother vehicles, suspending agents, antimicrobial preservatives,antioxidants and viscosity enhancing agents not listed in Table 7.Formulations may also contain additional ingredients that are notvehicles, suspending agents, antimicrobial preservatives, antioxidantsor viscosity enhancing agents. In some embodiments, the formulationsdescribed herein are intranasal formulations.

TABLE 7 Use Ingredients Vehicle Water, distilled water, purified water,water for injection, ethanol, buffers Suspending agent Alginic acid,bentonite, calcium stearate, carbomers, carboxymethylcellulose calcium,carboxymethylcellulose sodium, carrageenan, cellulose, ceratonia,colloidal silicon dioxide, dextrin, gelatin, guar gum, hydroxyethylcellulose, hydroxyethylmethyl cellulose, hydroxypropyl cellulose,hypromellose, kaolin, magnesium aluminum silicate, maltitol,medium-chain triglycerides, methylcellulose, microcrystalline cellulose,polycarbophil, polyethylene glycol, potassium alginate, povidone,propylene glycol alginate, sesame oil, sodium alginate, sodium starchglycolate, sorbitan esters, sucrose, tragacanth, xanthan gumAntimicrobial Alcohol, benzalkonium chloride, benzethonium chloride,benzoic acid, preservative benzyl alcohol, boric acid, bronopol,butylated hydroxyanisole, butylparaben, cetrimide, cetylpyridiniumchloride, chlorbutanol, chlorhexidine, chlorobutanol, chlorocresol,chloroform, chloroxylenol, cresol, dimethyl ether, ethylparaben,glycerin, hexetidine, imidurea, isopropyl alcohol, lactic acid,methylparaben, monothioglycerol, parabens, alkyl chain length, phenol,phenoxyethanol, phenylethyl alcohol, phenylmercuric acetate,phenylmercuric borate, phenylmercurie nitrate, potassium benzoate,potassium metabisulfite, potassium sorbate, propionic acid, propylgallate, propylene glycol, propylparaben, sodium acetate, sodiumbenzoate, sodium borate, sodium lactate, sodium metabisulfite, sodiumpropionate, sodium sulfite, sorbic acid, thimerosal, xylitol AntioxidantAscorbyl palmitate, butylated hydroxyanisole, butylated hydroxytoluene,malic acid, propyl gallate, sodium bisulfite, sodium sulfite, sodiummetabisulfite, potassium metabisulfite, potassium bisulfite, sodiumthiosulfate, sodium formaldehyde sulfoxylate, L-ascorbic acid,D-ascorbic acid, acetylcysteine, cysteine, thioglycerol, thioglycoUicacid, thiolactic acid, thiourea, dithiothreitol, dithioerythreitol,glutathione, nordihydroguaiaretic acid, tocopherol, sodium ascorbate,hypophophorous acid, fumaric acid Viscosity enhancing Acacia, agar,alginic acid, bentonite, carbomers, carboxymethylcellulose agentcalcium, carboxymethylcellulose sodium, carrageenan, ceratonia,ectostearyl alcohol, chitosan, colloidal silicon dioxide,cyclomethicone, ethylcelluose, gelatin, glycerin, glyceryl behenate,guar gum, hectorite, hydrogenated vegetable oil type I, hydroxyethylcellulose, hydroxyethylmethyl cellulose, hydroxypropyl cellulose,hydroxypropyl starch, hypromellose, magnesium aluminum silicate,maltodextrin, methylcellulose, polydextrose, polyethylene glycol,poly(methylvinyl ether/maleic anhydride), polyvinyl acetate phthalate,polyvinyl alcohol, potassium chloride, povidone, propylene glycolalginate, saponite, sodium alginate, sodium chloride, stearyl alcohol,sucrose, tragacanth, xanthan gum

Exemplary formulations (e.g., intranasal formulation) are shown in Table8.

TABLE 8 Formulation 1 Formulation 2 Formulation 3 Formulation 4Formulation 5 Vehicle Water for Water for Water for pH 6 Buffered Waterfor injection injection injection water for injection injectionSuspending None None None None Microcrystalline agent celluloseAntimicrobial None Benzoic acid Benzalkonium Benzoic acid Benzoic acidpreservative chloride Antioxidant None Thioglycerol Acetyl cysteineThioglycerol Ascorbic acid Viscosity None None None None Carboxymethylenhancing cellulose agent Other pH modifying pH modifying None pHmodifying pH modifying ingredients agents agents agents agents

Kits

Also provided are kits containing materials useful for the treatment ofa condition that is responsive to fulvestrant (e.g., cancer or systemiclupus erythematosus). The kits may contain an inclusion complex offulvestrant and a cyclodextrin, and optionally contain instructions foruse (e.g., instructions for preparation and/or administration of aformulation comprising the complex). Information detailing possible sideeffects of the formulation, and any other relevant information may alsobe enclosed. The instructions may be in any suitable format, including,but not limited to, printed matter, videotape, computer readable disk,optical disc or directions to internet-based instructions.

In one aspect, is provided a kit for treating an individual who suffersfrom or is susceptible to one or more of the conditions describedherein, comprising a first container comprising a dosage amount of aformulation containing an inclusion complex as disclosed herein, andinstructions for use. The container may be any of those known in the artand appropriate for storage and delivery of intravenous, intranasal ororal formulations. In certain embodiments the kit further comprises asecond container comprising a pharmaceutically acceptable carrier,diluent, adjuvant, etc. for preparation of the formulation to beadministered to the individual.

In some embodiments, the kits comprise a container with a label.Suitable containers include, for example, bottles, vials, and testtubes. The containers may be formed from a variety of materials such asglass or plastic. The containers may hold an inclusion complexcomprising fulvestrant and a cyclodextrin. In some embodiments, thecontainers may further comprise one or more additional pharmaceuticalagents (e.g., an anti-cancer agent). The label on the container mayindicate that the inclusion complex or the formulation is used fortreating or suppressing a condition that is responsive to fulvestrant(e.g., cancer or systemic lupus erythematosus), and may also indicatedirections for in vivo use, such as those described herein.

The kit may further include other materials desirable from a commercialand user standpoint, including other buffers, diluents, filters,needles, syringes, and package inserts with instructions for performingany methods described herein. In some embodiments, the kit comprises thecontainer described above and a second container comprising a buffer.

The kits may include additional pharmaceutical agents for use inconjunction with the formulation described herein. In one aspect, theadditional pharmaceutical agent is fulvestrant in uncomplexed form, suchas in the product FASLODEX® (AstraZeneca Pharmaceuticals LP). In somevariations, the additional pharmaceutical agent(s) may be one or moredrug(s) for the treatment of conditions responsive to fulvestrant (e.g.,cancer or systemic lupus erythematosus). In some variations, theadditional pharmaceutical agent(s) may be one or more drug(s) for thetreatment of one or more side effects from the use of the inclusioncomplexes described herein. These agents may be provided in a separateform, or mixed with the complexes described herein, provided such mixingdoes not reduce the effectiveness of either the pharmaceutical agent orformulation described herein and is compatible with the route ofadministration. Similarly the kits may include additional agents foradjunctive therapy or other agents known to the skilled artisan aseffective in the treatment or prevention of the conditions describedherein.

In some embodiments, the additional pharmaceutical agent is ananti-cancer agent. In some embodiments, the anti-cancer agent is anaromatase inhibitor (AI), including, but not limited to, anastrozole(e.g., ARIMIDEX®), letrozole (e.g., FEMARA®), exemestane (e.g.,AROMASIN®), aminoglutethimide (Cytadren®), vorozole (e.g., RIVIZOR®),formestane (e.g., LENTARON®), fadrozole (e.g. AFEMA®), or testolactone(e.g., TESLAC®). In some embodiments, the anti-cancer agent is anepidermal growth factor receptor (EGFR) inhibitor, including, but notlimited to, gefitinib (e.g., IRESSA®), trastuzumab (e.g., HERCEPTIN®),or erlotinib (e.g., TARCEVA®). In some embodiments, the anti-canceragent is a farnesyl transferase inhibitor, including, but not limited totipifarnib (e.g., ZARNESTRA®). In some embodiments, the anti-canceragent is uncomplexed fulvestrant, such as uncomplexed fulvestrantadministered as an intramuscular injection (e.g., FASLODEX®). In someembodiments, the anti-cancer agent is an antioxidant, including, but notlimited to vitamin E, vitamin C, beta-carotene, and selenium.

In some embodiments, the anti-cancer agent is selected from the groupconsisting of anastrozole (e.g., ARIMIDEX®), letrozole (e.g., FEMARA®),exemestane (e.g., AROMASIN®), aminoglutethimide (e.g., Cytadren®),vorozole (e.g., RIVIZOR®), formestane (e.g., LENTARON®), fadrozole(e.g., AFEMA®), testolactone (e.g., TESLAC®), gefitinib (e.g., IRESSA®),trastuzumab (e.g., HERCEPTIN®), erlotinib (e.g., TARCEVA®), tipifarnib(e.g., ZARNESTRA®), uncomplexed fulvestrant (e.g., FASLODEX®), and anantioxidant (e.g., vitamin E, vitamin C, beta-carotene, and selenium).

Kits may also be provided that contain sufficient dosages of thecompounds described herein (including formulations thereof) to provideeffective treatment for an individual, such as a human in need of suchtreatment, for an extended period, such as any of 1-3 days, 1-5 days, aweek, 2 weeks, 3, weeks, 4 weeks, 6 weeks, 8 weeks, 3 months, 4 months,5 months, 6 months, 7 months, 8 months, 9 months or more.

The kits may include the composition as described herein packaged ineither a unit dosage form or in a multi-use form. The kits may alsoinclude multiple units of the unit dose form. The kits may be used forany of the methods described herein, including, for example, to treat anindividual with a condition described herein, or to delay a conditiondescribed herein. In certain embodiments the kits may include a dosageamount of at least one formulation as disclosed herein. Kits may alsocomprise a means for the delivery of the formulation thereof, such as asyringe, inhaler, intranasal spray, patch or other such device.

Methods of Use

The inclusion complexes described herein comprising fulvestrant and acyclodextrin, formulations thereof, and formulations comprising anuncomplexed fulvestrant and a cyclodextrin, or mixtures thereof, may beused for the treatment of indications that are believed to be or areresponsive to fulvestrant therapy (e.g., cancer and systemic lupuserythematosus). In some embodiments, provided is a method of treating acancer in an individual in need thereof, comprising administering to theindividual an effective amount of an inclusion complex comprisingfulvestrant and a cyclodextrin, or a formulation thereof.

For purposes herein, beneficial or desired results in a treatmentinclude, but are not limited to, one or more of the following:decreasing one or more symptoms resulting from the condition (e.g.,cancer or systemic lupus erythematosus), diminishing the extent of thedisease, stabilizing the condition, delaying or slowing the progressionof the condition, reversing the progression or severity of thecondition, ameliorating the condition, decreasing the dose of one ormore other medications required to treat the condition, and/orincreasing the quality of life of an individual who has been or issuspected of having the condition. The methods described hereincontemplate any one or more of these aspects of treatment. In oneaspect, a method of delaying the development of a condition is provided,wherein the method reduces the probability of developing the conditionin a given time frame and/or reduces the extent of the condition in agiven time-frame, when compared to not using the method. Suchcomparisons may be based on clinical studies, using a statisticallysignificant number of subjects. In a particular aspect, a method ofdelaying the development of a condition is provided, wherein the methodencompasses preventing a recurrence of the condition in a given timeframe, such as 3 months, 6 months, 1 year, 2 years, 3 years, 5 years, 10years or more.

In some embodiments, the methods and/or inclusion complex formulationsused herein reduce the severity of one or more symptoms associated withthe condition (e.g., cancer or systemic lupus erythematosus) by at leastany of about 10%, 20%, 30%, 400/%, 50%, 60%, 70%, 80%, 90%, 95%, or 100%compared to the corresponding symptom in the same individual prior totreatment or compared to the corresponding symptom in other individualsnot receiving the methods and/or inclusion complex formulations.

In some variations, the individual being treated for a conditiondescribed herein (e.g., cancer or systemic lupus erythematosus) has beenidentified as having one or more of the symptoms described herein.Identification of the conditions as described herein by a skilledphysician is routine in the art such as routine physical exams orclinical detection.

In some embodiments of a method directed to cancer, the cancer is earlystage cancer, non-metastatic cancer, primary cancer, advanced cancer,locally advanced cancer, metastatic cancer, cancer in remission,recurrent cancer, cancer in an adjuvant setting, cancer in a neoadjuvantsetting, or cancer substantially refractory to hormone therapy. In someembodiments, the cancer includes, but is not limited to, breast cancer,endometrial cancer, prostate cancer, and lung cancer.

The methods detailed herein may be applicable to an individual in needthereof, which may be an individual who has been diagnosed with,previously treated for, and/or suspected of having the condition to betreated (e.g., cancer or systemic lupus erythematosus). With respect toprevention, the individual in need thereof may also be an individual whois at risk for a condition (e.g., age, a family or self history of thecondition, life-style factors indicative of risk for the condition,etc.). In a particular variation, an individual is an individual who haspreviously had the condition and is at risk fir recurrence of thecondition.

In some embodiments, the individual is a mammal, including, but notlimited to human, bovine, horse, feline, rabbit, canine, rodent, orprimate. In some embodiments, the mammal is a primate. In someembodiments, the primate is a human. In some embodiments, the individualis human, including adults and children. The human may be of either sexand the methods may be applicable to either a man or a woman. In oneaspect, the individual is a human who is believed to be or is in need oftreatment. In some embodiments, the individual is a pre-menopausal,per-menopausal or post-menopausal woman. In some embodiments, theindividual is a post-menopausal woman with disease (e.g., breast cancer)progression following hormone (e.g., antiestrogen) therapy. In someembodiments, the individual is a pre-menopausal woman with disease(e.g., breast cancer) progression following hormone (e.g.,anti-estrogen) therapy. In some embodiments, the individual is anon-mammal. In some variations, the primate is a non-human primate suchas chimpanzees and other apes and monkey species. In some embodiments,the mammal is a farm animal such as cattle, horses, sheep, goats, andswine; pets such as rabbits, dogs, and cats; laboratory animalsincluding rodents, such as rats, mice, and guinea pigs; and the like. Insome embodiments, the individual is a non-mammal, including, but notlimited to, birds, and the like. The term “individual” does not denote aparticular age or sex.

In some embodiments, provided is a method of treating breast cancer inan individual in need thereof, comprising administering to theindividual an effective amount of an inclusion complex comprisingfulvestrant and a cyclodextrin (e.g., MBCD or HPBCD), or a formulationthereof. In some embodiments, the individual is a pre- orperi-menopausal individual with breast cancer progression followinganti-estrogen therapy. In some embodiments, the individual is apost-menopausal individual with breast cancer progression followinganti-estrogen therapy. In some embodiments, the breast cancer is earlystage breast cancer, non-metastatic breast cancer, advanced breastcancer, stage IV breast cancer, locally advanced breast cancer,metastatic breast cancer, hormone receptor positive metastatic breastcancer, advanced breast cancer, breast cancer in remission, breastcancer in an adjuvant setting, ductal carcinoma in situ (DCIS), invasiveductal carcinoma (IDC), or breast cancer in a neoadjuvant setting. Insome specific embodiments, the breast cancer is hormone receptorpositive metastatic breast cancer. In some embodiments, the breastcancer is advanced breast cancer. In some embodiments, the breast canceris ductal carcinoma in situ. In some embodiments, there are providedmethods of treating breast cancer (which may be HER2 positive or HER2negative), including, for example, advanced breast cancer, stage IVbreast cancer, locally advanced breast cancer, and metastatic breastcancer. In some embodiments, the individual may be a human who has agene, geneticmutation, or polymorphism associated with breast cancer(e.g., BRCA1, BRCA2, ATM, CHEK2, RAD51, AR, DIRAS3, ERBB2, TP53, AKT,PTEN, and/or PI3K) or has one or more extra copies of a gene (e.g., oneor more extra copies of the HER2 gene) associated with breast cancer. Insome embodiments, the method further comprises identifying a cancerpatient population (e.g., breast cancer population) based on a hormonereceptor status of patients having tumor tissue not expressing both ERand PgR and administering to the patient population an effective amountof the inclusion complex comprising fulvestrant and a cyclodextrin.

In some embodiments, provided is a method of treating endometrial cancer(or uterine cancer) in an individual in need thereof, comprisingadministering to the individual an effective amount of an inclusioncomplex comprising fulvestrant and a cyclodextrin (e.g., MBCD or HPBCD),or a formulation thereof. In some embodiments, the individual is a pre-or peri-menopausal woman. In some embodiments, the individual is apost-menopausal woman. In some embodiments, the endometrial cancer isendometrial stromal sarcoma, uterine carcinosarcoma, endometrialadenocarcinoma, Type I endometrial carcinoma, or Type II endometrialcarcinoma. In some embodiments, the endometrial carcinoma is stage IA,stage IB, stage IC, stage IIA, stage IIB, stage IIIA, stage IIB, stageIIIC, stage IVA, or stage IVB endometrial carcinoma.

In some embodiments, provided is a method of treating prostate cancer inan individual in need thereof, comprising administering to theindividual an effective amount of an inclusion complex comprisingfulvestrant and a cyclodextrin (e.g., MBCD or HPBCD), or a formulationthereof. In some embodiments, the prostate cancer is an adenocarcinoma.In some embodiments, the prostate cancer is a sarcoma, neuroendocrinetumor, small cell cancer, ductal cancer, or a lymphoma. There areprovided methods of treating prostate cancer at any of the four stages,A, B, C, or D, according to the Jewett staging system. In someembodiments, the prostate cancer is stage A prostate cancer (the cancercannot be felt during a rectal exam.). In some embodiments, the prostatecancer is stage B prostate cancer (the tumor involves more tissue withinthe prostate, it can be felt during a rectal exam, or it is found with abiopsy that is done because of a high PSA level.). In some embodiments,the prostate cancer is stage C prostate cancer (the cancer has spreadoutside the prostate to nearby tissues.). In some embodiments, theprostate cancer is stage D prostate cancer. In some embodiments, theprostate cancer may be androgen independent prostate cancer (AIPC). Insome embodiments, the prostate cancer may be androgen dependent prostatecancer. In some embodiments, the prostate cancer may be refractory tohormone therapy. In some embodiments, the prostate cancer may besubstantially refractory to hormone therapy. In some embodiments, theindividual may be a human who has a gene, genetic mutation, orpolymorphism associated with prostate cancer (e.g., RNASEL/HPC1,ELAC2/HPC2, SR-A/MSR1, CHEK2, BRCA2, PON1, OGG1, MIC-1, TLR4, and/orPTEN) or has one or more extra copies of a gene associated with prostatecancer.

In some embodiments, provided is a method of treating lung cancer in anindividual in need thereof, comprising administering to the individualan effective amount of an inclusion complex comprising fulvestrant and acyclodextrin (e.g., MBCD or HPBCD), or a formulation thereof. In someembodiments, the hmg cancer is a non-small cell lung cancer (NSCLC).Examples of NSCLC include, but are not limited to, large-cell carcinoma(e.g., large-cell neuroendocrine carcinoma, combined large-cellneuroendocrine carcinoma, basaloid carcinoma, lymphoepithelioma-likecarcinoma, clear cell carcinoma, and large-cell carcinoma with rhabdoidphenotype), adenocarcinoma (e.g., acinar, papillary (e.g.,bronchioloalveolar carcinoma, nonmucinous, mucinous, mixed mucinous andnonmucinous and indeterminate cell type), solid adenocarcinoma withmucin, adenocarcinoma with mixed subtypes, well-differentiated fetaladenocarcinoma, mucinous (colloid) adenocarcinoma, mucinouscystadenocarcinoma, signet ring adenocarcinoma, and clear celladenocarcinoma), neuroendocrine lung tumors, and squamous cell carcinoma(e.g., papillary, clear cell, small cell, and basaloid). In someembodiments, the NSCLC may be, according to TNM classifications, a stageT tumor (primary tumor), a stage N tumor (regional lymph nodes), or astage M tumor (distant metastasis). In some embodiments, the lung canceris a carcinoid (typical or atypical), adenosquamous carcinoma,cylindroma, or carcinoma of the salivary gland (e.g., adenoid cysticcarcinoma or mucoepidermoid carcinoma). In some embodiments, the lungcancer is a carcinoma with pleomorphic, sarcomatoid, or sarcomatouselements (e.g., carcinomas with spindle and/or giant cells, spindle cellcarcinoma, giant cell carcinoma, carcinosarcoma, or pulmonary blastoma).In some embodiments, the cancer is small cell lung cancer (SCLC; alsocalled oat cell carcinoma). The small cell lung cancer may belimited-stage, extensive stage or recurrent small cell lung cancer. Insome embodiments, the individual may be a human who has a gene, geneticmutation, or polymorphism suspected or shown to be associated with lungcancer (e.g., SASH1, LATS1, IGF2R, PARK2, KRAS, PTEN, Kras2, Krag, Pas1,ERCC1, XPD, IL8RA, EGFR, a_(f)AD, EPHX, MMP1, MMP2, MMP3, MMP12,IL1p\RAS, and/or AKT) or has one or more extra copies of a geneassociated with lung cancer.

In another aspect, the inclusion complexes described herein (e.g., aninclusion complex comprising fulvestrant and a cyclodextrin) may be usedfor the treatment of systemic lupus erythematosus. In some embodimentsis provided a method of treating systemic lupus erythematosus in anindividual, comprising administering to the individual an effectiveamount of an inclusion complex comprising fulvestrant and acyclodextrin, or a formulation thereof.

The method detailed herein may use any inclusion complex or compositiondetailed herein. As described below, the inclusion complex may beadministered via any route (e.g., orally, intranasally, sublingually,parenterally, such as intravenously, rectally, intravaginally).

In another aspect, provided is a method of treating a cancer in anindividual in need thereof, comprising administering to the individualan effective amount of an inclusion complex comprising fulvestrant and acyclodextrin, or a formulation thereof, and one other pharmaceuticalagent.

In another aspect, provided is a method of treating systemic lupuserythematosus in an individual in need thereof, comprising administeringto the individual an effective amount of an inclusion complex comprisingfulvestrant and a cyclodextrin, or a formulation thereof and one otherpharmaceutical agent.

In the context of an inclusion complex of fulvestrant, the additionalpharmaceutical agent in one aspect refers to an active agent other thancomplexed fulvestrant, for example, an anti-cancer agent other thanfulvestrant, which is administered to elicit a therapeutic effect orfulvestrant in an uncomplexed form. In one aspect, the additionalpharmaceutical agent is an anti-cancer agent other than fulvestrant.More than one additional pharmaceutical agent may be employed. Anadditional pharmaceutical agent may be directed to a therapeutic effectrelated to one or more conditions that fulvestrant is intended to treator prevent (e.g., cancer or systemic lupus erythematosus) or thepharmaceutical agent may be intended to treat or prevent a symptom ofthe underlying condition (e.g., pain, swelling, warmth and rednessthroughout the breast, Paget's disease of the breast, malar rash,fatigue, loss of appetite, inflammation (e.g., pericarditis,myocarditis, or endocarditis), bone or joint pain, arthritis, anemia andiron deficiency, pleuritis, pleural effusion, lupus pneumonitis,pulmonary hypertension, pulmonary emboli, pulmonary hemorrhage, chronicdiffuse interstitial lung disease, hematuria, headache, or proteinuria,etc.) or to further reduce the appearance or severity of side effects offulvestrant (e.g., asthenia, pain, headache, back pain, vasodilatation,nausea, vomiting, constipation, anemia, bone pain, arthritis, dizziness,insomnia, rash, or urinary tract infection).

In some embodiments, the pharmaceutical agent is an anti-cancer agent.In some embodiments, the anti-cancer agent is an aromatase inhibitor(AI), including, but not limited to, anastrozole (e.g., ARIMIDEX®),letrozole (e.g., FEMARA®), exemestane (e.g., AROMASIN®),aminoglutethimide (Cytadren®), vorozole (e.g., RIVIZOR®), formestane(e.g., LENTARON®), fadrozole (e.g., AFEMA®), or testolactone (e.g.,TESLAC®). In some embodiments, the anti-cancer agent is an epidermalgrowth factor receptor (EGFR) inhibitor, including, but not limited to,gefitinib (e.g., IRESSA®), trastuzumab (e.g., HERCEPTIN®), or erlotinib(e.g., TARCEVA®). In some embodiments, the anti-cancer agent is afarnesyl transferase inhibitor, including, but not limited to tipifamib(e.g., ZARNESTRA®). In some embodiments, the anti-cancer agent isfulvestrant in uncomplexed form (e.g., FASLODEX®). In some embodiments,the anti-cancer agent is an antioxidant, including, but not limited tovitamin E, vitamin C, beta-carotene, and selenium. In some embodiments,the formulation may include combinations of two or more of theanti-cancer agents as described herein (e.g., any of 2, 3, or moreanti-cancer agents).

In some embodiments, the antioxidant includes, but is not limited to,ascorbyl palmitate, butylated hydroxyanisole, butylated hydroxytoluene,malic acid, propyl gallate, sodium bisulfite, sodium sulfite, sodiummetabisulfite, potassium metabisulfite, potassium bisulfite, sodiumthiosulfate, sodium formaldehyde sulfoxylate, L-ascorbic acid,D-ascorbic acid, acetylcysteine, cysteine, thioglycerol, thioglycoUicacid, thiolactic acid, thiourea, dithiothreitol, dithiocrythreitol,glutathione, nordihydroguaiaretic acid, tocopherol, sodium ascorbate,hypophophorous acid, and fumaric acid.

In some embodiments, the anti-cancer agent is selected from the groupconsisting of anastrozole (e.g., ARIMIDEX®), letrozole (e.g., FEMARA®),exemestane (e.g., AROMASIN®), aminoglutethimide (Cytadren®), vorozole(e.g., RIVIZOR®), formestane (e.g., LENTARON®), fadrozole (e.g.,AFEMA®), testolactone (e.g., TESLAC®), gefitinib (e.g., IRESSA®),trastuzumab (e.g., HERCEPTIN®), erlotinib (e.g., TARCEVA®), tipifarnib(e.g., ZARNESTRA®), fulvestrant in uncomplexed form (e.g., FASLODEX®),and an antioxidant (e.g., vitamin E, vitamin C, beta-carotene, andselenium).

In some embodiments, provided is a method of treating a hormone receptorpositive metatstatic breast cancer in an individual in need thereof,comprising administering to the individual an effective amount of aninclusion complex comprising fulvestrant and a cyclodextrin, or aformulation thereof, and anastrozole.

In some embodiments, provided is a method of treating a hormone receptorpositive metatstatic breast cancer in an individual in need thereofcomprising administering to the individual an effective amount of aninclusion complex comprising fulvestrant and a cyclodextrin, or aformulation thereof, and exemestane.

In some embodiments, provided is a method of treating a hormone receptorpositive metatstatic breast cancer in an individual in need thereof,comprising administering to the individual an effective amount of aninclusion complex comprising fulvestrant and a cyclodextrin, or aformulation thereof, and uncomplexed fulvestrant (e.g., FASLODEX®).

In some embodiments, provided is a method of treating a hormone receptorpositive metatstatic breast cancer in an individual in need thereofcomprising administering to the individual an effective amount of aninclusion complex comprising fulvestrant and a cyclodextrin, or aformulation thereof and gefitinib.

In some embodiments, provided is a method of treating a hormone receptorpositive metatstatic breast cancer in an individual in need thereofcomprising administering to the individual an effective amount of aninclusion complex comprising fulvestrant and a cyclodextrin, or aformulation thereof and lapatinib.

In some embodiments, provided is a method of treating a hormone receptorpositive metatstatic breast cancer in an individual in need thereof,comprising administering to the individual an effective amount of aninclusion complex comprising fulvestrant and a cyclodextrin, or aformulation thereof and tipifarnib.

In some embodiments, the inclusion complex comprising fulvestrant and acyclodextrin, or a formulation thereof, and the other pharmaceuticalagent are administered simultaneously. In some embodiments, theinclusion complex comprising fulvestrant and a cyclodextrin, or aformulation thereof, and the other pharmaceutical agent are administeredsequentially. Either the inclusion complex or the other pharmaceuticalagent may be administered first. The inclusion complex and the otherpharmaceutical agent may be contained in the same or different packagesor compositions.

Dosing and Methods of Administration

The amount of the inclusion complex administered to an individual (suchas a human) and/or the amount administered in order to achieve aneffective amount may vary based on a variety of factors, including, forexample, the particular condition being treated, the frequency ofadministration, the particular formulation being administered, theseverity of the condition being treated and the age, weight and generalhealth of the individual, the adverse effects experienced by theindividual being treated, etc. A pharmaceutical unit dosage chosen maybe fabricated and administered to provide a defined final concentrationof drug in the blood, tissues, organs, or other targeted region of thebody. Determination of an effective amount for a given situation can bereadily determined by routine experimentation (e.g., using in vivoanimal models) and is within the skill and judgment of the ordinaryclinician, particularly in view of the teachings provided herein.

In various embodiments, an effective amount of the inclusion complex ortherapy may do any one or more of the following: (i) reduce the numberof cancer cells; (ii) reduce tumor size; (iii) inhibit, retard, slow tosome extent, and preferably stop cancer cell infiltration intoperipheral organs; (iv) inhibit (e.g., slow to some extent andpreferably stop) tumor metastasis; (v) inhibit tumor growth; (vi)prevent or delay occurrence and/or recurrence of a tumor, and (vii)relieve to some extent one or more of the symptoms associated with thecancer. In various embodiments, the amount is sufficient to ameliorate,palliate, lessen, and/or delay one or more of symptoms of cancer. Invarious embodiments, an effective amount of the inclusion complex ortherapy may inhibit, retard, slow to some extent, prevent, delayoccurrence and/or recurrence of; or relieve to some extent one or moreof the symptoms associated with systemic lupus erythematosus.

The effective amount may vary depending on the composition beingadministered, the condition being treated/prevented, the severity of thecondition being treated/prevented, the age, body size, weight, andrelative health of the individual, the route and form of administration,the judgement of the attending medical or veterinary practitioner (ifapplicable), and other factors appreciated by the skilled artisan inview of the teaching provided herein. An effective amount may beassessed, for example, by using data from one or more clinical,physiological, biochemical, histological, electrophysiological, and/orbehavioral evaluations.

In some embodiments, the amount of the inclusion complex comprisingfulvestrant and a cyclodextrin is effective to result in an objectiveresponse (such as a partial response or a complete response). In someembodiments, the amount of inclusion complex is sufficient to result ina complete response in the individual. In some embodiments, the amountof the inclusion complex is sufficient to result in a partial responsein the individual. In some embodiments, the amount of the inclusioncomplex administered alone is sufficient to produce an overall responserate of more than about any of about 10%, 20%, 30%, 40%, 50%, 60%, 70%,80%, 90%, or 95% among a population of individuals treated with thecomplex. Responses of an individual to treatment can be determined bythe skilled artisan using, for example routine physical exams and/orclinical detection known in the art and/or described herein.

In some embodiments, the amount of the inclusion complex is below thelevel that induces a toxicological effect (where the toxicologicaleffect in one aspect is an effect above a clinically acceptable level oftoxicity) or is at a level where a potential side effect can becontrolled or tolerated when the complex is administered to theindividual. In some embodiments, the amount of the inclusion complex isclose to a maximum tolerated dose (MTD) of the complex following thesame dosing regime. In some embodiments, the amount of the inclusioncomplex is more than any of about 80%, 90%, 95%, or 98% of the MTD.

In some embodiments, the amount of fulvestrant from an inclusion complexcomprising fulvestrant and a cyclodextrin is included in any of thefollowing ranges: about 0.1 to about 5 mg, about 1 to about 10 mg, about5 to about 10 mg, about 10 to about 15 mg, about 15 to about 20 mg,about 20 to about 25 mg, about 20 to about 50 mg, about 25 to about 50mg, about 50 to about 75 mg, about 50 to about 100 mg, about 75 to about100 mg, about 100 to about 125 mg, about 125 to about 150 mg, about 150to about 175 mg, about 175 to about 200 mg, about 200 to about 225 mg,about 225 to about 250 mg, about 250 to about 300 mg, about 300 to about350 mg, about 350 to about 400 mg, about 400 to about 450 mg, about 450to about 500 mg, about 550 to about 600 mg, about 650 to about 700 mg,or about 750 to about 800 mg. In some embodiments, the amount offulvestrant from an inclusion complex is in the range of about 1 mg toabout 800 mg, such as about 5 mg to about 700 mg, 10 mg to about 500 mg,20 mg to about 250 mg, or about 25 mg, 50 mg, 75 mg, 100 mg, 125 mg, 150mg, 175 mg, 200 mg, 250 mg, 300 mg, 350 mg, 400 mg, 500 mg, 600 mg, 700mg, or 800 mg. In some embodiments of the liquid formulations, theconcentration of fulvestrant from an inclusion complex comprisingfulvestrant and cyclodextrin as inclusion complex is dilute (about 0.1mg/ml) or concentrated (about 200 mg/ml), including for example any ofabout 0.1 to about 200 mg/ml, about 0.1 to about 100 mg/ml, about 1 toabout 50 mg/ml, about 2 mg/ml to about 25 mg/ml, about 4 to about 10mg/ml, about 5 mg/ml. In some embodiments, the concentration offulvestrant of the inclusion complex is at least about any of 0.5 mg/ml,1.3 mg/ml, 1.5 mg/ml, 2 mg/ml, 3 mg/ml, 4 mg/ml, 5 mg/ml, 6 mg/ml, 7mg/ml, 8 mg/ml, 9 mg/ml, 10 mg/ml, 15 mg/ml, 20 mg/ml, 25 mg/ml, 30mg/ml, 40 mg/ml, 50 mg/ml, 60 mg/ml, 70 mg/ml, 80 mg/ml, 90 mg/ml, 100mg/ml, 150 mg/ml, or 200 mg/ml.

Exemplary effective amounts of fulvestrant from an inclusion complexcomprising fulvestrant and a cyclodextrin include, but are not limitedto, any of about 2.5 mg/m², 5 mg/m², 10 mg/m², 15 mg/m², 20 mg/m², 25mg/m², 30 mg/m², 50 mg/m², 60 mg/m², 75 mg/m², 80 mg/m², 90 mg/m², 100mg/m², 120 mg/m², 160 mg/m², 175 mg/m², 180 mg/m², 200 mg/m², 210 mg/m²,220 mg/m², 250 mg/m², 260 mg/m², 300 mg/m², 350 mg/m², 400 mg/m², 500mg/m², 540 mg/m², 750 mg/m², 1000 mg/m², or 1080 mg/m². In variousvariations, a formulation includes less than about any of 350 mg/m², 300mg/m², 250 mg/m², 200 mg/m², 150 mg/m², 120 mg/m², 100 mg/m², 90 mg/m²,50 mg/m², or 30 mg/m² of fulvestrant. In some embodiments, the amount offulvestrant within the inclusion complex per administration is less thanany of about 25 m/m, 22 m/m², 20 mg/m², 18 mg/m², 15 mg/m², 14 mg/m², 13mg/m², 12 mg/m², 11 mg/m², 10 mg/m², 9 mg/m², 8 mg/m², 7 mg/m², 6 mg/m²,5 mg/m², 4 mg/m², 3 mg/m², 2 mg/m², or 1 mg/m. In some embodiments, theeffective amount of fulvestrant from the inclusion complex is includedin any of the following ranges: about 1 to about 5 mg/m, about 5 toabout 10 mg/m², about 10 to about 25 mg/m², about 25 to about 50 mg/m²,about 50 to about 75 mg/m², about 75 to about 100 mg/m², about 100 toabout 125 mg/m² about 125 to about 156 mg/m², about 150 to about 175mg/m², about 175 to about 200 mg/m², about 200 to about 225 mg/m², about225 to about 250 mg/m², about 250 to about 300 mg/m², about 300 to about350 mg/m², or about 350 to about 400 mg/m².

In some embodiments of any of the above aspects, the effective amount offulvestrant from an inclusion complex comprising fulvestrant and acyclodextrin includes at least any of about 0.01 mg/kg, 0.05 mg/kg, 0.1mg/kg, 0.25 mg/kg, 0.5 mg/kg, 1 mg/kg, 1.5 mg/kg, 2 mg/kg, 2.5 mg/kg,3.5 mg/kg, 5 mg/kg, 6.5 mg/kg, 7.5 mg/kg, 10 mg/kg, 15 mg/kg, or 20mg/kg. In various variations, the effective amount of fulvestrantincludes less than any of about 350 mg/kg, 300 mg/kg, 250 mg/kg, 200mg/kg, 150 mg/kg, 100 mg/kg, 50 mg/kg, 25 mg/kg, 20 mg/kg, 10 mg/kg, 7.5mg/kg, 6.5 mg/kg, 5 mg/kg, 3.5 mg/kg, 2.5 mg/kg, 2 mg/kg, 1.5 mg/kg, 1mg/kg, 0.75 mg/kg, 0.5 mg/kg, 0.25 mg/kg, 0.1 mg/kg, 0.05 mg/kg.

Exemplary dosing frequencies include, but are not limited to, weeklywithout break; weekly, three out of four weeks; once every three weeks;once every two weeks; weekly, two out of three weeks. In someembodiments, the inclusion complex is administered any of about onceevery 2 weeks, once every 3 weeks, once every 4 weeks, once every 6weeks, or once every 8 weeks. In some embodiments, the composition isadministered at least any of about 1×, 2×, 3×, 4×, 5×, 6×, or 7× (i.e.,daily) a week. In some embodiments, the intervals between eachadministration are less than any of about 6 months, 3 months, 1 month,20 days, 15, days, 12 days, 10 days, 9 days, 8 days, 7 days, 6 days, 5days, 4 days, 3 days, 2 days, or 1 day. In some embodiments, theintervals between each administration are more than any of about 1month, 2 months, 3 months, 4 months, 5 months, 6 months, 8 months, or 12months. In some embodiments, there is no break in the dosing schedule.In some embodiments, the interval between each administration is no morethan about a week. The administration of the inclusion complex can beextended over an extended period of time, such as from about a month upto about seven years. In some embodiments, the composition isadministered over a period of at least any of about 2, 3, 4, 5, 6, 7, 8,9, 10, 11, 12, 18, 24, 30, 36, 48, 60, 72, or 84 months. The dosingfrequency of the inclusion complex may be adjusted over the course ofthe treatment based on the judgment of the administering physician. Forexample, a multiple daily dosage routine of the inclusion complex may beincluded.

The inclusion complexes described herein allow, in some embodiments,infusion of the complex to an individual over an infusion time that isshorter than about 24 hours. For example, in some embodiments, theinclusion complex is administered over an infusion period of less thanany of about 24 hours, 12 hours, 8 hours, 5 hours, 3 hours, 2 hours, 1hour, 30 minutes, 20 minutes, or 10 minutes. In some embodiments, theinclusion complex is administered over an infusion period of about 30minutes.

Any of the inclusion complexes described herein (e.g., an inclusioncomplex comprising fulvestrant and a cyclodextrin) can be administeredto an individual (such as human) via various routes, including, forexample, intravenous, intra-arterial, intraperitoneal, intrapulmonary,oral, inhalation, intravesicular, intramuscular, intra-tracheal,subcutaneous, intraocular, intrathecal, transmucosal, intranasal,rectal, intravaginal and transdermal. In some embodiments, sustainedcontinuous release formulation of the composition may be used. In onevariation, inclusion complex is administered by any acceptable routeincluding, but not limited to, orally, intramuscularly, transdermally,intravenously, intranasally, rectally, intravaginally through an inhaleror other air borne delivery systems and the like. Additional methods ofadministration are known in the art.

In some embodiments, the inclusion complexes described herein (e.g., aninclusion complex comprising fulvestrant cyclodextrin) are administeredparenterally (e.g., intravenously). For example, in some embodiments areprovided methods of treating a cancer in an individual (e.g., an adult)comprising intranasally administering an inclusion complex comprisingfulvestrant and a cyclodextrin.

The physiochemical properties of the inclusion complexes describedherein (e.g., an inclusion complex comprising fulvestrant and acyclodextrin) may allow the complexes to be taken orally orsublingually. In some embodiments, the inclusion complexes orformulations comprising the complexes are suitable for oral orsublingual administration.

As described herein, the inclusion complexes may be administered with anadditional therapeutic agent and/or an additional treatment modality.The dosing frequency of the inclusion complex and the additionalpharmaceutical agent may be adjusted over the course of the treatmentbased on the judgment of the administering physician. When administeredseparately, the inclusion complex and the additional therapeutic agentcan be administered at different dosing frequency or intervals. Forexample, the inclusion complex can be administered weekly, while theadditional therapeutic agent can be administered more or lessfrequently. In some embodiments, sustained continuous releaseformulation of the inclusion complex and/or the additional therapeuticagent may be used. Various formulations and devices for achievingsustained release are known in the art. A combination of theadministration configurations described herein can be used.

In some embodiments, the inclusion complex (e.g., the inclusion complexcomprising fulvestrant and a cyclodextrin) can be administered daily andthe additional therapeutic agent (e.g., FASLODEX®) can be administeredmonthly. In some embodiments, the inclusion complex (e.g., the inclusioncomplex comprising fulvestrant and a cyclodextrin) can be administeredweekly and the additional therapeutic agent (e.g., FASLODEX®) can beadministered monthly.

In some embodiments of any of the aspects of the inventions providedherein, the cyclodextrin is hydroxypropyl beta-cyclodextrin (HPBCD) ormethyl beta-cyclodextrin (MBCD). In some embodiments, the cyclodextrinis hydroxypropyl beta-cyclodextrin (HPBCD) or methyl beta-cyclodextrin(MBCD), wherein HPBCD or MBCD is used in a formulation for intravascularadministration. In some embodiments, the cyclodextrin is hydroxypropylbeta-cyclodextrin (HPBCD) or methyl beta-cyclodextrin (MBCD), whereinHPBCD or MBCD is used in a formulation for slow intravascularadministration.

The commercial product (e.g., FASLODEX®) is to be stored underrefrigerated conditions (2-8° C.), whilst the inventor has found thatexcellent stability at room temperature may be attained by the inclusioncomplex of the present invention: an inclusion complex of the inventionhas been found to be stable at temperatures of over 25° C. and at 40° C.for over 24 months.

The present invention will be understood more readily by reference tothe following examples, which are provided by way of illustration andare not intended to be limiting of the present invention.

EXAMPLES

It is understood that the examples and embodiments described herein arefor illustrative purposes only and that various modifications or changesin light thereof will be suggested to persons skilled in the art and areto be included within the spirit and purview of this application.

The fulvestrant used in the Examples was obtained from Hangzhou HysenPharma Co., Ltd., Hangzhou, China.

Example 1: Evaluation of Cyclodextrins for Fulvestrant AqueousSolubility Enhancement

A comprehensive selection of pharmaceutically acceptable cyclodextrinswas selected for the solubility enhancement evaluation with fulvestrant.The most relevant characteristics of these cyclodextrins are presentedin Table 9.

TABLE 9 Characteristics summary of cyclodextrins used in fulvestrantaqueous solubility evaluation. Water Number of Solubility GlucopyranoseMolecular (25° C.) Cyclodextrin Type Trade Name Units AbbreviationWeight (mg/mL) Alpha-cyclodextrin CAVAMAX ® 6 ACD 973 145 W6 PharmaBeta-cyclodextrin CAVAMAX ® 7 BCD 1135 18.5 W7 Pharma Methyl beta-CAVASOL ® 7 MBCD 1310 >750 cyclodextrin W7 M Pharma HydroxypropylKLEPTOSE ® 7 HPBCD 1100 >750 beta-cyclodextrin HPB PharmaSulfobutylether 7 SBEBCD 2163 >500 beta-cyclodextrin Gamma- CAVAMAX ® 8GCD 1297 232 cyclodextrin W8 Pharma Hydroxypropyl CAVASOL ® 8 HPGCD1574 >750 gamma-cyclodextrin W8 HP Pharma

HPLC Instrumentation and Chemicals

The HPLC system used for the chromatographic determination offulvestrant consisted of a Waters 600E quaternary gradient pump, aWaters 717plus WISP autosampler and a Waters 2487 Dual A, Absorbancedetector. Separation was performed using a Phenomenex Luna C18 column(250×4.6 mm; 5 urn) (Phenomenex, Torrance, Calif.) (supplied bySeparations, South Africa) and a mobile phase consisting of 50%MeCN:22.5% ddH₂0:27.5% MeOH. The mobile phase was degassed and filteredprior to use. An injection volume of 50 uL was selected, together with aflow rate of 1.2 mL/min, which resulted in fulvestrant peak retentiontimes of approximately 11.2 minutes. Sample run times were 15 minutes.The UV detector wavelength was set to 280 nm.

The fulvestrant used for preparation of standard solutions, was ofpharmaceutical grade. The acetonitrile used as mobile phase was of HPLCgrade (Merck Chemicals, South Africa). Double distilled water (ddH₂0)was used at all times during the assay.

Stock fulvestrant solution was prepared according to the followingsteps: 1) 10 mg of fulvestrant powder was accurately weighed into a 10mL volumetric flask; 2) the fulvestrant powder was dissolved and made tovolume in mobile phase; 3) this solution was then vortexed for at least20 seconds to ensure complete dissolution of the analyte. Aconcentration range of 10-500 μg/mL fulvestrant was prepared in themobile phase. Calibration data generated during the fulvestrant aqueoussolubility study for one of the fulvestrant calibration plots ispresented in Table 10, with the graphical presentation of the datapresented in FIG. 1. The method, which produced a linear plot, issuitable for the evaluation of fulvestrant.

TABLE 10 Fulvestrant HPLC calibration data. Fulvestrant Concentration(μg/mL) Peak Area 10 6017.79 20 11356.84 50 29814.88 100 57946.34 200114870.10 300 173177.22 400 229736.60 500 287091.62

Results

a. Alpha-Cyclodextrin (ACD)

The aqueous solubility of fulvestrant was evaluated in the presence ofincreasing concentrations of ACD and the absence thereof.

1 mL of each of the following aqueous solutions was prepared containing0; 25; 50; 75 and 100 mg/mL ACD in ddHiO. Excess fulvestrant was addedto each of these solutions and the samples were shaken for 24 hours onan orbital shaker at 200 rpm and at 22° C. room temperature. Excessfulvestrant was present in all samples at all times. After 24-hours, thesamples were filtered through 0.45 μm syringe tip filters, dilutedaccording to requirements and assayed by HPLC. The fulvestrant aqueoussolubility results are presented in Table 11 and FIG. 2.

TABLE 11 Fulvestrant aqueous solubility data in the presence and absenceof various concentrations of ACD. Quantity of Quantity of ACDFulvestrant Fulvestrant Average True Fulvestrant Sample ConcentrationDiluted (μg/mL) Diluted (μg/mL) Concentration Concentration No. (mg/mL)Run 1 Run 2 (μg/mL) (mg/mL) 1 0 <10.000 <10.010 <10.00 <0.010 2 2533.620 32.970 33.30 0.033 3 50 110.600 110.880 110.74 0.111 4 75 310.460310.170 310.32 0.310 5 100 162.660 173.850 168.26 1.683b. Beta-Cyclodextrin (BCD)

The aqueous solubility of fulvestrant was evaluated in the presence ofincreasing concentrations of BCD and the absence thereof.

1 mL of each of the following aqueous solutions was prepared containing0; 5; 7.5; 10 and 15 mg/mL BCD in ddH₂O. Excess fulvestrant was added toeach of these solutions and the samples were shaken for 24 hours on anorbital shaker at 200 rpm and at 22° C. room temperature. Excessfulvestrant was present in all samples at all times. After 24-hours, thesamples were filtered through 0.45 μm syringe tip filters, dilutedaccording to requirements and assayed by HPLC. The fulvestrant aqeuoussolubility results are presented in Table 12 and FIG. 3.

TABLE 12 Fulvestrant aqueous solubility data in the presence and absenceof various concentrations of BCD. Quantity of Quantity of BCDFulvestrant Fulvestrant Average True Fulvestrant Sample ConcentrationDiluted (μg/mL) Diluted (μg/mL) Concentration Concentration No. (mg/mL)Run 1 Run 2 (μg/ml) (mg/mL) 1 0 <10.000 <10.000 <10.00 <0.010 2 5<10.000 <10.000 <10.00 <0.010 3 7.5 <10.000 <10.000 <10.00 <0.010 4 10<10.000 <10.000 <10.00 <0.010 5 15 13.060 12.940 13.00 0.013c. Methyl Beta-Cyclodextrin (MBCD)

The aqueous solubility of fulvestrant was evaluated in the presence ofincreasing concentrations of MBCD and the absence thereof.

1 mL of each of the following aqueous solutions was prepared containing0; 50; 100; 150; 200; 250; 300; 350 and 400 mg/mL MBCD in ddH₂0. Excessfulvestrant was added to each of these solutions and the samples wereshaken for 24 hours on an orbital shaker at 200 rpm and at 22° C. roomtemperature. Excess fulvestrant was present in all samples at all times.After 24-hours, the samples were filtered through 0.45 μm syringe tipfilters, diluted according to requirements and assayed by HPLC. Thefulvestrant aqueous solubility results are presented in Table 13 andFIG. 4.

TABLE 13 Fulvestrant aqueous solubility data in the presence and absenceof various concentrations of MBCD. Quantity of Quantity of MBCDFulvestrant Fulvestrant Average True Fulvestrant Sample ConcentrationDiluted (μg/mL) Diluted (μg/mL) Concentration Concentration No. (mg/mL)Runl Run 2 (μg/mL) (mg/mL) 1 0 <10.00 <10.00 <10.00 <0.010 2 50 74.6175.37 74.99 0.750 3 100 302.12 301.99 302.06 3.021 4 150 67.40 67.4767.44 6.744 5 200 106.87 107.01 106.94 10.694 6 250 174.30 174.28 174.2917.429 7 300 226.52 226.94 226.73 22.673 8 350 293.21 292.29 292.7529.275 9 400 359.13 358.71 358.92 35.892d. Hydroxypropyl Beta-Cyclodextrin (HPBCD)

The aqueous solubility of fulvestrant was evaluated in the presence ofincreasing concentrations of HPBCD and the absence thereof.

1 mL of each of the following aqueous solutions was prepared containing0; 50; 100; 150; 200; 250; 300; 350 and 400 mg/mL HPBCD in ddH₂0. Excessfulvestrant was added to each of these solutions and the samples wereshaken for 24 hours on an orbital shaker at 200 rpm and at 22° C. roomtemperature. Excess fulvestrant was present in all samples at all times.After 24-hours, the samples were filtered through 0.45 μm syringe tipfilters, diluted according to requirements and assayed by HPLC. Thefulvestrant aqueous solubility results are presented in Table 14 andFIG. 5.

TABLE 14 Fulvestrant aqueous solubility data in the presence and absenceof various concentrations of HPBCD. Quantity of Quantity of HPBCDFulvestrant Fulvestrant Average True Fulvestrant Sample ConcentrationDiluted (μg/mL) Diluted (μg/mL) Concentration Concentration No. (mg/mL)Runl Run 2 (μg/mL) (mg/mL) 1 0 <10.00 <10.00 <10.00 <0.010 2 50 21.0920.81 20.95 0.210 3 100 48.32 48.27 48.30 0.483 4 150 93.82 93.50 93.660.937 5 200 156.30 42.30 99.30 1.563 6 250 118.71 118.97 118.84 2.377 7300 164.70 166.54 165.62 3.312 8 350 215.41 215.10 215.26 4.305 9 400275.20 272.14 273.67 5.473e. Sulfobutylether Beta-Cyclodextrin (SBEBCD)

The aqueous solubility of fulvestrant was evaluated in the presence ofincreasing concentrations of SBEBCD and the absence thereof.

1 mL of each of the following aqueous solutions was prepared containing0; 50; 100; 150; 200; 250; 300; 350 and 400 mg/mL SBEBCD in ddH₂0.Excess fulvestrant was added to each of these solutions and the sampleswere shaken for 24 hours on an orbital shaker at 200 rpm and at 22° C.room temperature. Excess fulvestrant was present in all samples at alltimes. After 24-hours, the samples were filtered through 0.45 μm syringetip filters, diluted according to requirements and assayed by HPLC. Thefulvestrant aqueous solubility results are presented in Table 15 andFIG. 6.

TABLE 15 Fulvestrant aqueous solubility data in the presence and absenceof various concentrations of SBEBCD. Quantity of Quantity of SBEBCDFulvestrant Fulvestrant Average True Fulvestrant Sample ConcentrationDiluted (μg/mL) Diluted (μg/mL) Concentration Concentration No. (mg/mL)Runl Run 2 (μg/mL) (mg/mL) 1 0 <10.00 <10.00 <10.00 <0.010 2 50 76.9877.61 77.30 0.077 3 100 167.7 166.49 167.10 0.167 4 150 273.01 272.83272.92 0.273 5 200 398.52 399.09 398.81 0.399 6 250 304.32 305.15 304.740.609 7 300 425.82 425.96 425.89 0.852 8 350 382.24 377.67 379.96 1.1409 400 503.13 501.21 502.17 1.507f. Gamma-Cyclodextrin (GCD)

The aqueous solubility of fulvestrant was evaluated in the presence ofincreasing concentrations of GCD and the absence thereof.

1 mL of each of the following aqueous solutions was prepared containing0; 50; 100; 150; and 200 mg/mL GCD in ddH₂0. Excess fulvestrant wasadded to each of these solutions and the samples were shaken for 24hours on an orbital shaker at 200 rpm and at 22° C. room temperature.Excess fulvestrant was present in all samples at all times. After24-hours, the samples were filtered through 0.45 μm syringe tip filters,diluted according to requirements and assayed by HPLC. The fulvestrantaqueous solubility results are presented in Table 16 and FIG. 7.

TABLE 16 Fulvestrant aqueous solubility data in the presence and absenceof various concentrations of GCD. Quantity of Quantity of GCDFulvestrant Fulvestrant Average True Fulvestrant Sample ConcentrationDiluted (μg/mL) Diluted (μg/mL) Concentration Concentration No. (mg/mL)Runl Run 2 (μg/mL) (mg/mL) 1 0 <10.000 <10.000 <10.00 <0.010 2 50 10.70010.530 10.62 0.011 3 100 <10.000 <10.000 <10.00 <0.010 4 150 <10.000<10.000 <10.00 <0.010 5 200 <10.000 <10.000 <10.00 <0.010g. Hydroxypropyl Gamma-Cyclodextrin (HPGCD)

The aqueous solubility of fulvestrant was evaluated in the presence ofincreasing concentrations of HPGCD and the absence thereof.

1 mL of each of the following aqueous solutions was prepared containing0; 50; 100; 150; 200; 250; 300; 350 and 400 mg/mL HPGCD in ddH₂0. Excessfulvestrant was added to each of these solutions and the samples wereshaken for 24 hours on an orbital shaker at 200 rpm and at 22° C. roomtemperature. Excess fulvestrant was present in all samples at all times.After 24-hours, the samples were filtered through 0.45 μm syringe tipfilters, diluted according to requirements and assayed by HPLC. Thefulvestrant aqueous solubility results are presented in Table 17 andFIG. 8.

TABLE 17 Fulvestrant aqueous solubility data in the presence and absenceof various concentrations of HPGCD. Quantity of Quantity of IIPGCDFulvestrant Fulvestrant Average True Fulvestrant Sample ConcentrationDiluted (μg/mL) Diluted (μg/mL) Concentration Concentration No. (mg/mL)Runl Run 2 (μg/mL) (mg/mL) 1 0 <10.00 <10.00 <10.00 <0.010 2 50 38.6637.97 38.32 0.037 3 100 116.80 115.74 116.27 0.107 4 150 220.67 220.94220.81 0.203 5 200 334.75 334.66 334.71 0.300 6 250 474.68 472.99 473.840.215 7 300 400.24 384.76 392.50 0.603 8 350 500.71 505.22 502.97 0.8029 400 486.39 450.47 468.43 1.061

Discussion

From the results presented in Tables 11 to 17 and FIGS. 2 to 8, it isclear that fulvestrant forms inclusion complexes with cyclodextrins togreater and lesser extents, depending on the specific cyclodextrin.Fulvestrant aqueous solubility enhancements with ACD; SBEBCD and HPGCDyielded fulvestrant aqueous solubilities of between 1 to 2 mg/mL, whichwas a significant enhancement over the aqueous solubility of fulvestrantin the absence of these cyclodextrins. The aqueous solubilityenhancements of fulvestrant with BCD and GCD were lower. The aqueoussolubility of fulvestrant in the presence of HPBCD was enhanced frombelow 0.010 mg/mL to 5.5 mg/mL in 400 mg/mL HPBCD. The greatest aqueoussolubility enhancement of fulvestrant was, however, achieved with MBCD,whereby the fulvestrant aqueous solubility was significantly enhancedfrom below 0.010 mg/mL to 35.9 mg/mL in 400 mg/mL MBCD.

Example 2: Oral Fulvestrant Absorption Study in Male Whistar Rats

The comparative oral absorption of two novel aqueous cyclodextrin basedformulations of fulvestrant versus an uncomplexed fulvestrant aqueoussuspension (without any cyclodextrins) following oral administrations of10 mg/kg by gavage to adult male Whistar rats was assessed.

Materials and Methods

The study was conducted using eighteen male adult Whistar rats. Each ratpartook in a single experiment. Fulvestrant was administered at a doseof 10 mg/kg by oral gavage to each animal, with blood concentrationsmeasured hourly over a 3-hour period.

The test articles were considered to be >99% pure and contained 5 mg/mLof fulvestrant. The vehicles used to prepare the two novel batchescomprised 400 mg/mL hydroxypropyl beta-cyclodextrin (HPBCD) (Batch 1)and 400 mg/mL methyl beta-cyclodextrin (MBCD) (Batch 2). The controlfulvestrant formulation comprised of a 5 mg/mL fulvestrant suspension inddH₂0 (Batch 3). All animals were fasted for approximately 24 hoursprior to dose administration. Water was available ad libitum. Pre-dose,and 1 h and 2 h post dose blood sampling was conducted by means of thetail snip procedure, whereby 0.5 mL was collected at each samplingpoint. Five minutes prior to the final (3 h) blood sampling point, ratswere anaesthetised with 0.5 ml 6% sodium pentobarbitone and 5 mL bloodwas collected. Prior to drug administration by means of gavage, each ofthe rats was anaesthetized with halothane gas, following which a gavagetube was inserted through the mouth and through which the varioussolutions were administered. 0.5 mL saline was administered through thegavage tubes immediately following drug administration.

The information for Experiments I to III are summarized in Tables 18 to20 respectively.

TABLE 18 Experiment I Information. Formulation Rat No. Dose Dose volumeRat Mass Fulvestrant Batch 1 1 10 mg/kg 0.58 mL 0.290 kg FulvestrantBatch 1 2 10 mg/kg 0.54 mL 0.270 kg Fulvestrant Batch 1 3 10 mg/kg 0.51mL 0.254 kg Fulvestrant Batch 1 4 10 mg/kg 0.62 mL 0.309 kg FulvestrantBatch 1 5 10 mg/kg 0.53 mL 0.264 kg Fulvestrant Batch 1 6 10 mg/kg 0.54mL 0.270 kg

TABLE 19 Experiment II Information Formulation Rat No. Dose Dose volumeRat Mass Fulvestrant Batch 2 1 10 mg/kg 0.52 mL 0.261 kg FulvestrantBatch 2 2 10 mg/kg 0.54 mL 0.271 kg Fulvestrant Batch 2 3 10 mg/kg 0.61mL 0.304 kg Fulvestrant Batch 2 4 10 mg/kg 0.57 mL 0.287 kg FulvestrantBatch 2 5 10 mg/kg 0.60 mL 0.300 kg Fulvestrant Batch 2 6 10 mg/kg 0.57mL 0.284 kg

TABLE 20 Experiment III Information. Formulation Rat No. Dose Dosevolume Rat Mass Fulvestrant Batch 3 1 10 mg/kg 0.56 mL 0.278 kgFulvestrant Batch 3 2 10 mg/kg 0.51 mL 0.253 kg Fulvestrant Batch 3 3 10mg/kg 0.54 mL 0.271 kg Fulvestrant Batch 3 4 10 mg/kg 0.53 mL 0.267 kgFulvestrant Batch 3 5 10 mg/kg 0.53 mL 0.265 kg Fulvestrant Batch 3 6 10mg/kg 0.57 mL 0.284 kg

Blood Sampling and Bioanalytical Methods

All of the animals that were dosed by means of oral gavage were bledpre-dose, and at 1 hour (h) and 2 h post dose, with blood samplinghaving been conducted by means of the tail snip procedure, whereby 0.5mL was collected at each sampling point. Five minutes prior to theterminal blood sampling point (3 h), rats were anaesthetised with 0.5 ml6% sodium pentobarbitone and 5 mL blood was collected from each, afterwhich the rats were sacrificed.

The tail snip blood was collected in 1 mL K3E (K₃EDTA) MINICOLLECT®(Greiner Bio-one; Lot 090319; Exp. 2010-08) blood collection tubes, withthe terminal samples having been collected in 9 mL K3E (K₃EDTA)VACUETTE® (Greiner Bio-one; Lot A090602D; Exp. 2010-12) blood collectiontubes. Following collection, the samples were placed on ice and thencentrifuged at 4° C. for 15 minutes. 0.2 mL and 2 mL plasma aliquotswere obtained from the tail snip and terminal sampling pointsrespectively. These samples were frozen (−60° C. to −80° C.) prior tobeing shipped on dry ice to a facility for analysis of fulvestrantconcentrations.

Bioanalytical Methodology and Calculation of Pharmacokinetic (P)Parameters

The test article, fulvestrant, was measured in adult Whistar rat plasmavia a liquid chromatography-mass spectroscopy (LC-MS) method, asdescribed below.

A sensitive and selective LC-MS/MS method was developed and used todetermine fulvestrant in rat plasma. Prior to assay, rat plasma sampleswere thawed at room temperature, briefly vortexed, after which 100 uLplasma was pipetted into 1.5 ml Eppendorf® Safe-Lock® microcentrifugetubes. Samples were then precipitated with methanol (200 uL) containingthe internal standard, doxepin (245 ng/mL). These samples were vortexedfor 30 seconds and centrifuged for 5 minutes at 15 000 rpm. Thesupernatant (200 uL) was transferred to 96 well plates and 5 uL wasinjected onto the HPLC column. Calibration standards were prepared inrat plasma (anticoagulant lithium heparinate), whereby a stock solutionsof fulvestrant in methanol was used to spike a pool of blank rat plasmawhich was serially diluted with blank rat plasma to attain the desiredconcentrations, spanning a range of 4.5 ng/mL to 1149 ng/mL (STD 1-STD9).

Chromatography was performed on a Phenomenex, Luna Phenyl Hexyl,(150×2.1 mm, 5 um) analytical column. The mobile phase consisted ofmethanol and 5 mM ammonium formate (80:20 v/v) and was delivered at aconstant flow rate of 0.3 mL/min for 7.5 minutes. An Agilent 1100 seriesautosampler injected 5 uL onto the HPLC column. The injection needle wasrinsed with water:methanol (50:50) for 10 seconds using the flush portwash station. Samples were cooled to ˜5° C. while awaiting injection.Detection was performed on an API 4000 mass spectrometer (ESI in thepositive ion mode, MRM) and the settings on the apparatus are summarizedin tables below.

TABLE 21A ESI settings Curtain gas 30 Collision gas 7 Ionspray voltage(V) 5500 Source temperature (° C.) 450 Gas 1 (psi) 30 Gas 2 (psi) 40

TABLE 21B MS/MS settings Fulvestrant Internal standard Q1 mass [M + II]+607.3 279.9 Q3 mass 589.2 107.2 Dwell time (ms) 150 150 Declusteringpotential (V) 76 61 Entrance potential (V) 10 10 Collision energy (V) 2331 Collision cell exit potential (V) 22 8 Scan type MRM MRM Polaritypositive positive Pause time (ms) 5 5

Results

a. Plasma Concentration-Time Profiles for Orally AdministeredFulvestrant

Individual fulvestrant plasma concentrations for Experiments I, II, andIII are presented in Tables 22 to 24. Individual and mean plasmafulvestrant concentration-time profiles following oral administration bygavage for Experiments I, II, and III are presented in FIGS. 9 to 12.

TABLE 22 Fulvestrant plasma concentration data generated for ExperimentI (HPBCD). Plasma concentration (ng/mL) 0 h 1 h 2 h 3 h Rat 1 0.00 30.9020.10 11.20 Rat 2 0.00 23.80 11.60 9.63 Rat 3* 0.00 608.00 239.00 45.70Rat 4 0.00 76.70 30.30 21.80 Rat 5 0.00 237.00 103.00 26.40 Rat 6 0.0062.00 135.00 20.50 *The data from rat 3 in Experiment I was regarded asbeing an outlier and as such, it was not included in the average datacalculation.

TABLE 23 Fulvestrant plasma concentration data generated for ExperimentII (MBCD). Plasma concentration (ng/mL) 0 h 1 h 2 h 3 h Rat 1 0.00 45.4058.20 15.50 Rat 2 0.00 72.10 98.90 21.30 Rat 3 0.00 121.00 71.60 14.00Rat 4 0.00 177.00 102.00 10.60 Rat 5 0.00 191.00 83.00 36.40 Rat 6 0.0068.90 61.60 18.80 Average 0.00 96.63 68.19 17.09

TABLE 24 Fulvestrant plasma concentration data generated for ExperimentIII (No CD). Plasma concentration (ng/ml) 0 h L h 2 h 3 h Rat 1 0.000.00 0.00 0.00 Rat 2 0.00 0.00 0.00 0.00 Rat 3 0.00 0.00 0.00 0.00 Rat 40.00 32.40 4.75 0.00 Rat 5 0.00 0.00 0.00 0.00 Rat 6 0.00 0.00 0.00 0.00Average 0.00 5.40 0.79 0.00

From the data generated during this study, it is clear that very littleto no oral absorption was observed for the control 5 mg/mL fulvestrantin ddH₂0 suspension, which is in agreement with published literature,indicating that oral administration of fulvestrant presents with verypoor bioavailability, thus rendering the compound unsuitable for oralformulation. In contrast to the control fulvestrant suspensionformulation, both of the cyclodextrin based fulvestrant formulations(HPBCD and MBCD) presented with oral absorption. The HPBCD and vMBCDbased formulations presented with similar absorption plots, as can beseen in FIG. 12, whereby the mean C_(max) values for the HPBCD and MBCDwere measured at 86.1 ng/mL and 96.6 ng/mL respectively and T_(max)occurring for both formulations at the 1 hour sampling point. The datafrom rat 3 in the HPBCD group was not included in the pooled data, asits C_(max) of 608.0 ng/mL is regarded as being an outlier

Comparison of the data generated following the administration by oralgavage for the two novel cyclodextrin based fulvestrant formulations andthe reference fulvestrant suspension formulation, revealed that verylimited to no oral absorption was observed for the control 5 mg/mLfulvestrant in WFI suspension, whereby low fulvestrant concentrationswere observed in two samples of rat 4 only. This observation is inagreement with published literature, indicating that oral administrationof fulvestrant presents with very poor bioavailability, thus renderingthe compound unsuitable for oral formulation. In contrast to the controlfulvestrant suspension formulation, both of the cyclodextrin basedfulvestrant formulations (HPBCD and MBCD) presented with fulvestrantbeing recovered from all post-dose samples collected from these twogroups of animals, thus indicating that the two cyclodextrin basedformulations do enhance oral bioavailability of fulvestrantsignificantly.

The HPBCD and MBCD based formulations presented with similar fulvestrantplasma concentrations over time, as may be evidenced by the datapresented in FIG. 12, whereby the mean C_(max) values for the HPBCD andMBCD formulations were measured at 86.1 ng/mL and 96.6 ng/mLrespectively and T_(max) occurring for both formulations at the 1 hoursampling point.

The data generated during the present study indicates that aqueousformulations of fulvestrant complexed with HPBCD or MBCD offer viableopportunities as oral formulations. Oral formulations present withdistinct advantages over the currently marketed long acting IMfulvestrant formulations, which present with pain upon injection, owingto the 5 mL injection volume. In addition to the formulation relatedside effects, long acting IM fulvestrant (250 mg/5 mL), takes 3-6 monthsto achieve steady state. A viable oral formulation can achieve thedesired plasma levels immediately, thus replacing the need for an IMformulation, or can be used together with the IM formulation as part ofa loading-dose regimen.

Example 3: Oral Fulvestrant Absorption Study Using Fulvestrant andHydroxypropyl Beta-Cyclodextrin (HPBCD) in Whistar Rats

Fulvestrant was administered at a dose of 5 mg/kg by oral gavage to eachanimal, with blood concentrations measured hourly over a 4-hour period.The vehicle used to prepare the novel batch comprised 450 mg/mLhydroxypropyl beta-cyclodextrin (HPBCD) (Batch 7_4). All animals werefasted for approximately 24 hours prior to dose administration. Waterwas available ad libitum. Pre-dose, and 1, 2 and 3 hour post dose bloodsampling for the oral gavage group was conducted by means of the tailsnip procedure, whereby 0.5 mL was collected at each sampling point.Five minutes prior to the final (4-hour) blood sampling point, rats wereanaesthetised with 0.5 ml 6% sodium pentobarbitone and 5 mL blood wascollected. Prior to drug administration by means of gavage, each of therats were anaesthetized with halothane gas, following which a gavagetube was inserted through the mouth and through which the varioussolutions were administered. 0.5 mL saline was administered through thegavage tubes immediately following drug administration. The informationfor this example is summarized in Table 25.

TABLE 25 Experiment information (HPBCD Oral) Rat Formulation No. SexDose Dose volume Rat Mass Fulvestrant Batch7_4 1 Male 5 mg/kg 0.305 mL0.305 kg Fulvestrant Batch7_4 2 Male 5 mg/kg 0.325 mL 0.325 kgFulvestrant Batch7_4 3 Male 5 mg/kg 0.350 mL 0.350 kg FulvestrantBatch7_4 4 Male 5 mg/kg 0.335 mL 0.335 kg Fulvestrant Batch7_4 5 Male 5mg/kg 0.310 mL 0.310 kg Fulvestrant Batch7_4 6 Male 5 mg/kg 0.274 mL0.274 kg

Blood Sampling and Bioanalytical Methods

The tail snip blood was collected in 1 mL K3E (K₃EDTA) MINICOLLECT®(Greiner Bio-one; Lot 090319; Exp. 2010-08) blood collection tubes, withthe terminal samples having been collected in 4 mL K3E (K₃EDTA)VACUETTE® (Greiner Bio-one; Lot L020902; Exp. 2010-08) blood collectiontubes. Following collection, these samples were placed on ice and thencentrifuged at 4° C. for 15 minutes. 0.2 mL and 2 mL plasma aliquotswere obtained from the tail-snip and terminal sampling pointsrespectively. These samples were frozen (−60° C. to −80° C.) prior tobeing shipped on dry ice to a facility for analysis of fulvestrantconcentrations. The test article, fulvestrant, was measured in adultWhistar rat plasma by via an LC-MS method. Individual and mean plasmafulvestrant concentration-time profiles following oral administration bygavage in this example are presented in FIG. 13 and Table 26.

TABLE 26 Fulvestrant plasma concentration (ng/mL) data generated fororal fulvestrant-HPBCD (5 mg/kg) Time (h) 0 1 2 3 4 Rat 1 0.0 33.6 24.325.6 15.4 Rat 2 0.0 87.5 65.0 37.0 9.2 Rat 3 0.0 22.1 14.3 22.0 16.1 Rat4 0.0 10.9 9.4 11.1 15.7 Rat 5 0.0 62.4 36.7 19.7 5.6 Rat 6 0.0 31.328.2 22.9 12.2 Average 0.0 41.3 29.7 23.1 12.4

Example 4: Oral Fulvestrant Dose-Ranging Absorption Study in Male andFemale Beagle Dogs

The purpose of the study was to investigate the pharmacokinetics of anovel fulvestrant-hydroxypropyl-beta-cyclodextrin (fulvestrant-HPBCD)solution following oral administration at various dose levels, to thatof a short acting (SA) fulvestrant intramuscular formulation in fastedmale and female beagle dogs (n=6).

Materials and Methods

The study was conducted using three male and three female adult beagledogs for a total sample size of n=6. Each dog participated in each ofthe five experiments. Experiments began with the lowest oral dose andprogressed to the highest oral dose, with the study concluding with theShort Acting (SA) IM formulation. Animals were fasted for 12 hours priorto each study with access to ad libitum water. A minimum washout-periodof 48-hours was allowed between experiments. Oral drug administrationcomprised the use of oro-gastric tubes for the administration of thefulvestrant-HPBCD solution (5 mg/mL fulvestrant), with the IMadministration into the buttock. The following dosages of fulvestrantwere administered for the various experiments:

-   -   1. Experiment 1: 2.5 mg/kg fulvestrant (5 mg/mL fulvestrant; 450        mg/mL HPBCD)    -   2. Experiment 2: 5 mg/kg fulvestrant (5 mg/mL fulvestrant; 450        mg/mL HPBCD)    -   3. Experiment 3: 10 mg/kg fulvestrant (5 mg/mL fulvestrant; 450        mg/mL HPBCD)    -   4. Experiment 4: 15 mg/kg fulvestrant (5 mg/mL fulvestrant; 450        mg/mL HPBCD)    -   5. Experiment 5: 2.5 mg/kg fulvestrant (Short Acting (SA) IM        fulvestrant injection)

The oral fulvestrant solutions contained 5 mg/mL of fulvestrant in 450mg/mL HPBCD. The fulvestrant SA IM control fulvestrant formulation was ashort acting formulation, developed, described and used by AstraZeneca(20 mg/mL fulvestrant).

Venous blood samples (1 ml) were obtained from each animal at pr-doseand the following times post-dose (in minutes): 15, 30, 45, 60, 75, 90,105, 120, 135, 150, 165, 180, 210, 240, 300, 360, 480, 720, 1080 and1440. Samples were collected via implanted vascular ports in the jugularveins of the dogs and into tubes containing K₂EDTA anticoagulant.Following collection, the blood samples were placed on ice and thencentrifuged for 15 minutes. The plasma was harvested and approximately0.5 mL aliquots were stored frozen (−18° C.) prior to being shipped ondry ice to a contract laboratory for the determination of fulvestrant inthe plasma samples. The same liquid chromatography-mass spectroscopy(LC-MS) analytical method was employed as that described in Example 3.

Results

Results generated during this study demonstrated that absorption,following oral ingestion, was rapid and followed a first order process,with fulvestrant present in all of the first post-dose blood samples(T=15 min). A plot of the mean fulvestrant plasma concentrations overtime is presented in FIG. 14 (Exp 1=2.5 mg/kg oral; Exp 2-5 mg/kg oral;Exp 3=10 mg/kg oral; Exp 4=15 mg/kg oral and Exp 5=2.5 mg/kg IM).

The consistency of the pharmacokinetic parameters, following oraladministration, over a six-fold dose range, suggests that thepharmacokinetics of fulvestrant-HPBCD is linear. These parametersinclude T_(max); absorption and elimination halftimes; clearance andmean residence times. As the pharmacokinetics derived from oraladministration did not differ statistically between the various doses, amore accurate estimation of the pharmacokinetics of the new fulvestrantformulation administered orally to dogs can be obtained by averaging andcomparing the pharmacokinetic parameters of each individual subject.Mean values for these parameters are: Absorption halftime—19 min;Estimated bioavailability—6.5% (relative to and assuming 100%bioavailability for the LA IM fulvestrant formulation); T_(max)≦74 min;MRT—258 min; elimination halftime—55 min.

Absorption from the gastro-intestinal tract occurs more rapidly thanfrom the site of IM injection, as evidenced by the prolonged meanabsorption halftime of the latter (77 min) and longer T_(max) (223 min).Mean elimination halftimes calculated from IM dosing were considerablylonger (467 min), as was the mean MRT (801 min), than those obtainedfollowing oral administration. Bioavailability for the IM administrationwas assumed to be complete. The C_(max) attained following the IMadministration at a dose of 2.5 mg/kg, was as great as those values thatwere achieved by the 10 mg/kg and 15 mg/kg oral doses. IM administrationresulted in a mean AUC_(0-t) that was significantly greater those valuesthat resulted from all oral doses.

Whilst the oral administration of fulvestrant-HPBCD presented with loworal bioavailability (approximately 6.5%), clinically effectiveconcentrations of fulvestrant are systemically absorbed, due to theabsorption halftime (20 min) being significantly shorter than theelimination halftime (55 minutes). During the study, systemicfulvestrant concentrations in excess of purported therapeutic levels(approximately 23 ng/mL) were achieved using the lowest administeredoral dose of 2.5 mg/kg. Simulations suggest that eight-hour dosing todogs (5 mg/kg) would achieve average concentrations (26 ng/mL) that arein excess of the proposed therapeutic levels. FIG. 15 illustrates asimulation of oral dosing to an “average” beagle dog, employing aregimen of 5 mg/kg every eight hours for 24 hours. It is postulated thatsimilar rapid absorption may be achieved in humans.

Example 5—Stability Batches and Data

In total, six stability batches of fulvestrant in the presence ofcyclodextrins were produced to evaluate different fulvestrantconcentrations with different cyclodextrins exposed to 2° C.; 25° C.;40° C. over 24-months.

The batch details are summarized in Table 27 below:

TABLE 27 Fulvestrant-HPBCD stability study batch summaries. FulvestrantBatch Complexation Storage Vials per Batch Conc. CD size Complexationtemperature volume storage Storage No. (mg/mL) CD Type Conc. (mL) Time(h) (° C.) (mL) condition Temperatures 1_1 5.0 HPBCD 400 40 24 RT 0.8 2525° C.; 40° C. 1_2 2.5 HPBCD 350 40 24 RT 0.8 25 25° C.; 40° C. 1_3 5.0HPBCD 400 60 24 RT 1.0 30 2° C. 1_4 2.5 HPBCD 350 60 24 RT 1.0 30 2° C.2_1 10 RAMEB 350 70 1.5 60° C. 1.0 23 2° C.; 25° C.; 40° C. 2_2 25 RAMEB350 70 1.5 60° C. 1.0 23 2° C.; 25° C.; 40° C. RT = Room temperature~25° C.

Each of the batches were produced, whereby the pre-weighed cyclodextrinquantity was dissolved in ˜80% final volume double distilled water atambient temperature (˜25° C.). Pre-weighed fulvestrant API was thenadded to the cyclodextrin solution with vigorous stirring. The stirringwas conducted for 24-hours for batches 1_1; 1_2; 13 and 1_4 underambient conditions. Complexation was optimize for batches 2_1 and 2_2,whereby complexation duration was reduced to 1.5 hours with heating to60° C. Following complexation, all batches were made to volume withdouble distilled water and filtered through 0.45 μm syringe filters,prior to being filled into storage vials. Samples were assayed via HPLCfor fulvestrant content following manufacture and at each evaluationpoint (3; 6; 9; 12; 18 and 24-months post manufacture). The HPLC methodused was also specific to the detection of the presence of the principlefulvestrant degradation product, fulvestrant sulphone. The HPLC methodemployed is the same as that described in Example 1. At each samplingpoint, three vials were selected for fulvestrant quantification;evaluation for the presence of fulvestrant sulphone and pHdetermination.

Summary plots for the average recoveries of the various batches andstorage conditions are presented in FIGS. 16-21.

HPLC analysis of all stability samples of all batches indicated that allevaluated samples are stable, with none falling outside the 90-110%fulvestrant recovery range during the 18-24-month evaluation period at2° C.; 25° C. and 40° C. There was no visible precipitation present, norany increase in the presence of the principle fulvestrant degradationproduct, fulvestrant sulphone.

The disclosures of all publications, patents, patent applications andother references referred to herein are hereby incorporated herein byreference in their entireties.

1. An inclusion complex comprising a) a hydroxypropyl gamma-, orhydroxypropyl beta-cyclodextrin; and b) a compound of the formula (I):

or a salt thereof or hydrate of the foregoing; and a C_(max) of at leastabout 2-times greater than administered uncomplexed compound of formula(I) in the same amount and under the same conditions; or bioavailabilityat least about 50% over the bioavailability of the uncomplexed compoundof formula (I).
 2. The inclusion complex of claim 1, wherein the molarratio of the compound of formula (I) to the cyclodextrin is from about1:1 to about 1:300.
 3. The inclusion complex of claim 1, wherein theinclusion complex increases the solubility of the compound of formula(I), in deionized water at 20° C., by at least about 1.5-fold comparedto the solubility of the compound of formula (I) in uncomplexed formunder the same conditions.
 4. The inclusion complex of claim 1, whereinthe inclusion complex increases the bioavailability of the compound offormula (I) by at least about or about 50% over the bioavailability ofthe compound of formula (I) in uncomplexed form.
 5. (canceled)
 6. Theinclusion complex of claim 1, further comprising a carrier, wherein thecarrier is selected from the group consisting of a solvent, a mixedsolvent, a complexing agent, a filler, a diluent, a granulating agent, adisintegrant, a lubricant, a glidant, a pH-modifier, a tonicitymodifier, an adjuvant, and a binder. 7-10. (canceled)
 11. The inclusioncomplex of claim 1, wherein the compound of formula (I) is fulvestrantsulphoxide A, fulvestrant sulphoxide B, or a mixture of fulvestrantsulphoxide A and fulvestrant sulphoxide B.
 12. A stable formulationcomprising

a) a hydroxypropyl gamma-, or hydroxypropyl beta-cyclodextrin; b) acompound of the formula (I) or a salt thereof or hydrate of theforegoing; and c) a pharmaceutically acceptable carrier. 13-21.(canceled)
 22. A substantially pure form of the inclusion complex ofclaim
 1. 23. A method of treating a cancer responsive to anti-estrogentherapy and/or ER-downregulation in an individual in need thereof,comprising administering intranasally, topically, or transdermally tothe individual an effective amount of the inclusion complex of claim 1.24. The method of claim 23, further comprising administeringsimultaneously or sequentially one or more anti-cancer agents, whereinthe mode of administration of the one or more anti-cancer agents is thesame or different. 25-44. (canceled)
 45. A method of producing theinclusion complex of claim 1, comprising the steps of: a. admixing thecompound of the formula (I) with the cyclodextrin; and b. adding asuitable amount of solvent, mixed solvent, or buffer to the mixture ofstep (a) and mixing until a suspension or solution is formed. 46-48.(canceled)
 49. A method for improving solubility of a compound of theformula (I):

or a salt, hydrate, or solvate thereof in water comprising complexingthe compound of the formula (I) with a cyclodextrin. 50-76. (canceled)